Tricyclic compounds as modulators of TNF-α synthesis and as PDE4 inhibitors

ABSTRACT

The present invention relates to chemical compounds of Formula (I) are as herein defined, pharmaceutical compositions, and methods of use in the treatment of conditions or disorders mediated by TNF-α or by PDE4, including but not limited to ulcerative colitis and Crohn&#39;s disease.

FIELD OF THE INVENTION

The present invention relates to chemical compounds useful as inhibitorsof the activity of tumor necrosis factor alpha (TNF-α) and as inhibitorsof phosphdiesterase IV (PDE4), synthetic methods for their manufacture,and methods for use of such compounds.

BACKGROUND OF THE INVENTION

Tumor necrosis factor-alpha (TNF-α), also referred to as TNF, DIF,TNF-alpha, TNFA, and TNFSF2, is a cell-associated cytokine that isprocessed from a 26 kd precursor form to a 17 kd soluble form. TNF-α hasbeen shown to be a primary mediator in humans and in animals ofinflammation, fever, and acute phase responses, similar to thoseobserved during acute infection and shock. Excess TNF-α has been shownto be lethal. There is now considerable evidence that blocking theeffects of TNF-α by use of soluble TNF receptor or with specificneutralizing antibodies can be beneficial in a variety of circumstancesincluding autoimmune diseases such as rheumetoid arthritis (RA),non-insulin dependent diabetes mellitus (NIDDM or Type II diabetes), andCrohn's disease.

Tumor necrosis factor (TNF) was first found in the serum of mice andrabbits infected with Bacillus Calmette-Guerin or injected withendotoxin, and was recognized on the basis of its cytotoxic, antitumorand metabolic properties. Many cell and tissues can produce TNF but itsproduction is largely accomplished by macrophages and monocytes.

TNF can have a positive effect on the host organism by stimulatingneutrophils and monocytes and by inhibiting the replication of viruses.Moreover, TNF-α activates the immune defenses against parasites and actsdirectly or indirectly as a mediator in immune reactions, inflammatoryprocesses, and other processes in the body, although the mechanisms bywhich it works have not yet been clarified in a number of cases. Theadministration of TNF-α can also be accompanied by harmful phenomenasuch as shock and tissue damage, which can be remedied by means ofantibodies against TNF-α.

TNF-α appears to be a mediator of cachexia which can occur inchronically invasive, for example, parasitic, diseases. TNF-α alsoappears to play a major part in the pathogenesis of shock caused by gramnegative bacteria, for example, endotoxic shock; TNF-α would also appearto be implicated in some if not all the effects of lipopolysaccharides.TNF-α has also been postulated to have a function in the tissue damagewhich occurs in inflammatory processes in the joints and other tissues,and in the lethality and morbidity of the graft-versus host reaction(GVHR, Transplant Rejection). A correlation has also been reportedbetween the concentration of TNF in the serum and the fatal outcome ofmeningococcal diseases.

The administration of TNF-α over a lengthy period causes a state ofanorexia and malnutrition which has symptoms similar to those ofcachexia, which accompany neoplastic and chronic infectious diseases.

A protein derived from the urine of fever patients has a TNF inhibitingactivity; the effect of this protein is presumed to be due to acompetitive mechanism at the level of the receptors (similar to theeffect of the interleukin 1 inhibitor).

Anti-TNF-α antibodies (cA2) are effective in treating patients withrheumatoid arthritis (RA), which discovery led to an increased interestin finding novel TNF-α inhibitors as possible potent drugs for RA.Rheumatoid arthritis is an autoimmune chronic inflammatory diseasecharacterized by irreversible pathological changes in the joints. Inaddition to RA, TNF-α antagonists may also be used in numerouspathological conditions and diseases. Some proofs indicating thebiological importance of TNF-α were obtained by in vivo experiments inmice, in which mouse genes for TNF-α or its receptor were inactivated.Such animals are resistant to collagen-induced arthritis and toendotoxin-caused shock. In animal assays where the TNF-α level wasincreased, a chronic inflammatory polyarthritis occurred and itspathological picture was alleviated by inhibitors of TNF-α production.The treatment of such inflammatory and pathological conditions usuallyincludes the application of non-steroid antiinflammatory drugs (NSAIDs)and, in more severe cases, gold salts, D-penicillinamine or methotrexateare administered. These drugs act symptomatically, but they do not stopthe pathological process.

Novel approaches in the therapy of rheumatoid arthritis are based upondrugs such as tenidap, leflunomide, cyclosporin, FK-506 and uponbiomolecules neutralizing the TNF-α action. Commercially availabletherapies include etanercept (Enbrel®, Immunex), a fusion protein of thesoluble TNF-α receptor, and infliximab (Remicade®, Centocor), a chimericmonoclonal human and mouse antibody. Additionally, etanercept andinfliximab are also registered for the therapy of Crohn's disease.

Compounds which inhibit the production of TNF-α are believed useful in awide variety of diseases and disorders through mechanism basedtherapeutic intervention. TNF-α inhibitors are believed useful fordiseases including but not limited to viral, alcoholic, or drug-inducedacute and fulminant hepatitis, hepatic steatosis, both alcoholic andnon-alcoholic, viral and non-viral hepatitis, hepatic cirrhosis,autoimmune hepatitis, chronic active hepatitis, Wilson's disease,myasthenia gravis, idiopathic sprue, autoimmune inflammatory boweldisease, ulcerative colitis, Crohn's disease, inflammatory boweldiseases, endocrine ophthalmopathy, Grave's disease, sarcoidosis,primary biliary cirrhosis, pancreatitis, nephritis, endotoxin shock,septic shock, haemodynamic shock, sepsis syndrome, post ischemicreperfusion injury, malaria, mycobacterial infection, meningitis,psoriasis, asthma, chronic obstructive pulmonary disease (COPD),eosinophilia, congestive heart failure, fibrotic diseases, cysticfibrosis, pulmonary fibrosis, hepatic fibrosis, renal fibrosis,cachexia, graft rejection, rejection by transplantation, cancer,diseases involving angiogenesis, autoimmune diseases, ankylosingspondylitis, autoimmune encephalomyelitis, autoimmune hematologicaldisorders, hemolytic anemia, aplastic anemia, pure red cell anemia,idiopathic thrombocytopenia, systemic lupus erythematosus (SLE),polychondritis, scleroderma, Wegener granulomatosis, dermatomyositis,Reiter's syndrome, non infection uveitis, autoimmune keratitis,keratoconjunctivitis sicca, vernal keratoconjunctivitis, interstitiallung fibrosis, psoriatic arthritis, psoriasis and other benign ormalignant proliferative skin diseases, atopic dermatitis, urticaria,neurodegenerative disorders, Parkinson's disease, Alzheimer's disease,acute and chronic multiple sclerosis, cancer, viral infection, humanimmunodeficiency virus (HIV), cachexia, thrombosis, skin inflammatorydiseases, osteoarthritis (OA), osteoporosis, RA, emphysema, chronicbronchiolitis, allergic rhinitis, radiation damage, hyperoxic alveolarinjury, periodontal disease, non-insulin dependent diabetes mellitus(Type II diabetes), and insulin dependent diabetes mellitus (Juvenile orType I diabetes).

Phosphodiesterases (PDEs) comprise a superfamily of enzymes responsiblefor the hydrolysis and inactivation of the second messengers cyclicadenosine monophosphate (CAMP) and cyclic guanosine monophosphate(cGMP). Eleven different PDE families have been identified to date (PDE1to PDE11) which differ in substrate preference, catalytic activity,sensitivity to endogenous activators and inhibitors, and encoding genes.

The PDE4 isoenzyme family exhibits a high affinity for cyclic AMP buthas weak affinity for cyclic GMP. Increased cyclic AMP levels caused byPDE4 inhibition are associated with the suppression of cell activationin a wide range of inflammatory and immune cells, including lymphocytes,macrophages, basophils, neutrophils, and eosinophils. Moreover, PDE4inhibition decreases the release of the cytokine Tumor NecrosisFactor-alpha (TNF-α).

In view of these physiological effects, PDE4 inhibitors of variedchemical structures have been recently disclosed for the treatment orprevention of chronic and acute inflammatory diseases and of otherpathological conditions, diseases and disorders known to be susceptibleto amelioration by inhibition of PDE4.

PDE4 are thought to be useful in the treatment and/or prophylaxis of avariety of diseases/conditions, especially inflammatory and/or allergicdiseases, in mammals such as humans, for example: asthma, chronicobstructive pulmonary disease (COPD) (e.g. chronic bronchitis and/oremphysema), atopic dermatitis, urticaria, allergic rhinitis, allergicconjunctivitis, vernal conjunctivitis, eosinophilic granuloma,psoriasis, rheumatoid arthritis, septic shock, ulcerative colitis,Crohn's disease, reperfusion injury of the myocardium and brain, chronicglomerulonephritis, endotoxic shock, adult respiratory distresssyndrome, multiple sclerosis, cognitive impairment (e.g. in aneurological disorder such as Alzheimer's disease), depression, or pain.Ulcerative colitis and/or Crohn's disease are collectively oftenreferred to as inflammatory bowel disease.

SUMMARY OF THE INVENTION

The present invention includes a compound of Formula (I):

or a salt thereof, wherein

-   A¹ is O or S;-   A² is O or S;-   U is N or C—(CH₂)_(u)R^(U);-   V is N or C—(CH₂)_(v)R^(V);-   W is N or C—(CH₂)_(w)R^(W);-   X is N or C—(CH₂)_(x)R^(X);-   Y is N or C—(CH₂)_(y)R^(Y);-   Z is N or C—(CH₂)_(z)R^(Z);-   R¹ is —(CH₂)_(q)R^(Q);-   R² is —(CH₂)_(s)R^(S);-   wherein    -   each of q, s, u, v, w, x, y, and z individually is 0, 1, 2, 3,        or 4;    -   each of R^(Q), R^(S), R^(U), R^(V), R^(W), R^(X), R^(Y), and        R^(Z) is independently selected from the group consisting of        -   alkyl;        -   alkyl substituted with one or more R^(ak);        -   alkenyl;        -   alkynyl;        -   aryl;        -   aryl substituted with one or more R^(a);        -   azido;        -   cyano;        -   cycloalkyl;        -   cycloalkyl substituted with one or more R^(c);        -   fused cycloalkylaryl substituted with one or more R^(f1);        -   fused arylcycloalkyl substituted with one or more R^(f2);        -   fused heterocyclyaryl substituted with one or more R^(f3);        -   fused arylheterocyclyl substituted with one or more R^(f4);        -   fused cycloalkylheteroaryl substituted with one or more            R^(f5);        -   fused heteroarylcycloalkyl substituted with one or more            R^(f6);        -   fused heterocyclylheteroaryl substituted with one or more            R^(f7);        -   fused heteroarylheterocyclyl substituted with one or more            R^(f8);        -   halogen;        -   haloalkyl;        -   heterocyclyl;        -   heterocyclyl substituted with one or more R^(hc);        -   heteroaryl;        -   heteroaryl substituted with one or more R^(ha);        -   hydrogen;        -   —NR³R⁴;        -   —C(O)NR³R⁴;        -   —C(O)R⁵;        -   —C(O)₂R⁶;        -   —S(O)_(j)R⁷;        -   —OR⁸; and        -   nitro;        -   wherein            -   each of R³ and R⁴ is independently selected from the                group consisting of H, acyl, alkyl, alkoxy, alkoxyalkyl,                alkylsulfonyl, aryl, cycloalkyl, heterocyclyl and                heteroaryl;            -   j is 0, 1, or 2;            -   each R⁵, R⁶, R⁷, and R⁸ is independently selected from                the group consisting of                -   hydrogen;                -   alkyl;                -   alkyl substituted with one or more R^(ak);                -   alkenyl;                -   alkynyl;                -   alkoxy;                -   aryl;                -   aryl substituted with one or more R^(a);                -   cycloalkyl;                -   cycloalkyl substituted with one or more R^(c);                -   halogen;                -   haloalkyl;                -   heterocyclyl;                -   heterocyclyl substituted with one or more R^(hc);                -   heteroaryl;                -   heteroaryl substituted with one or more R^(ha);                    wherein each R^(a), R^(c), R^(f1), R^(f2), R^(f3),                    R^(f4), R^(f5), R^(f6), R^(f7), R^(hc) and R^(ha) is                    independently selected from the group consisting of                    acyl, alkyl, alkenyl, alkynyl, alkoxy, amide, amino,                    aryl, cyano, cycloalkyl, halogen, haloalkyl,                    haloalkoxy, heteroaryl, hydroxy, nitro, —C(O)OR⁹,                    —SO₂R¹⁰, —SR¹¹, —C(O)R¹², —C(O)NR¹³, —NH—SO₂—R¹⁴,                    —SO₂—NR¹⁵R¹⁶, and —SO₂—CH₂—SO₂—CH₃; and wherein                -    each R⁹, R¹⁰, R¹¹, R¹², R¹³ R¹⁴, R¹⁵, and R¹⁶ is                    independently selected from the group consisting of                    hydrogen, alkyl, alkenyl, alkynyl, aryl, cycloalkyl,                    heterocyclyl, and heteroaryl,-   provided that when R¹ is hydrogen, R^(S) is selected from the group    consisting of    -   alkyl;    -   alkyl substituted with one or more R^(ak);    -   alkenyl;    -   alkynyl;    -   aryl;    -   aryl substituted with one or more R^(a);    -   azido;    -   cyano;    -   cycloalkyl;    -   cycloalkyl substituted with one or more R^(c);    -   fused cycloalkylaryl substituted with one or more R^(f1);    -   fused arylcycloalkyl substituted with one or more R^(f2);    -   fused heterocyclyaryl substituted with one or more R^(f3);    -   fused arylheterocyclyl substituted with one or more R^(f4);    -   fused cycloalkylheteroaryl substituted with one or more R^(f5);    -   fused heteroarylcycloalkyl substituted with one or more R^(f6);    -   fused heterocyclylheteroaryl substituted with one or more        R^(f7);    -   fused heteroarylheterocyclyl substituted with one or more        R^(f8);    -   heterocyclyl;    -   heterocyclyl substituted with one or more R^(hc);    -   heteroaryl;    -   heteroaryl substituted with one or more R^(ha);    -   —NR³R⁴;    -   —C(O)NR³R⁴;    -   —C(O)R⁵;    -   —C(O)₂R⁶;    -   —S(O)_(j)R⁷;    -   —OR⁸; and    -   nitro.

The present invention also provides pharmaceutical compositionscomprising a compound of Formula (I) or a salt thereof.

In another embodiment, the present invention provides methods for thepreparation of pharmaceutical compositions comprising a compound ofFormula (I) or a salt thereof. The pharmaceutical compositions mayfurther comprise a pharmaceutically acceptable carrier, excipient,diluent, or mixture thereof.

In another embodiment, the present invention provides methods for theuse of a compound of Formula (I), or a salt thereof, and for the use ofpharmaceutical compositions comprising a compound of Formula (I) or asalt thereof. The compounds and pharmaceutical compositions of thepresent invention may be used for the treatment of human or animaldisorders.

Additional features of the present invention will be describedhereinafter. It is to be understood that the invention is not limited inits application to the details set forth in the foregoing or followingdescription but is capable of other embodiments and of being practicedor carried out in various ways.

DETAILED DESCRIPTION OF THE INVENTION

The following definitions are meant to clarify, but not limit, the termsdefined. If a particular term used herein is not specificially defined,such term should not be considered indefinite. Rather, terms are usedwithin their accepted meanings.

As used herein the term “alkyl” refers to a straight or branched chainhydrocarbon having one to twelve carbon atoms, which may be optionallysubstituted as herein further described, with multiple degrees ofsubstitution being allowed. Examples of “alkyl” as used herein include,but are not limited to, methyl, ethyl, propyl, isopropyl, isobutyl,n-butyl, tert-butyl, isopentyl, and n-pentyl.

As used throughout this specification, the number of atoms, such ascarbon atoms in an alkyl group, for example, will be represented by thephrase “C_(x)-C_(y) alkyl,” which refers to an alkyl group, as hereindefined, containing the from x to y, inclusive, carbon atoms. Similarterminology will apply for other terms and ranges as well. Oneembodiment of the present invention includes so-called ‘lower’ alkylchains of one to six carbon atoms. Thus, C₁-C₆ alkyl represents a loweralkyl chain as hereabove described.

As used herein the term “alkenyl” refers to a straight or branched chainaliphatic hydrocarbon having two to twelve carbon atoms and containingone or more carbon-to-carbon double bonds, which may be optionallysubstituted as herein further described, with multiple degrees ofsubstitution being allowed. Examples of “alkenyl” as used hereininclude, but are not limited to, vinyl, and allyl.

As used herein the term “alkynyl” refers to a straight or branched chainaliphatic hydrocarbon having two to twelve carbon atoms and containingone or more carbon-to-carbon triple bonds, which may be optionallysubstituted as herein further described, with multiple degrees ofsubstitution being allowed. An example of “alkynyl” as used hereinincludes, but is not limited to, ethynyl.

As used herein, the term “alkylene” refers to a straight or branchedchain divalent hydrocarbon radical having from one to ten carbon atoms,which may be optionally substituted as herein further described, withmultiple degrees of substitution being allowed. Examples of “alkylene”as used herein include, but are not limited to, methylene, ethylene,n-propylene, and n-butylene.

As used herein, the term “alkenylene” refers to a straight or branchedchain divalent hydrocarbon radical having from two to ten carbon atomsand containing one or more carbon-to-carbon double bonds, which may beoptionally substituted as herein further described, with multipledegrees of substitution being allowed. Examples of “alkenylene” as usedherein include, but are not limited to, vinylene, allylene, and2-propenylene.

As used herein, the term “alkynylene” refers to a straight or branchedchain divalent hydrocarbon radical having from two to ten carbon atomsand containing one or more carbon-to-carbon triple bonds, which may beoptionally substituted as herein further described, with multipledegrees of substitution being allowed. An example of “alkynylene” asused herein includes, but is not limited to, ethynylene.

As used herein, the term “cycloalkyl” refers to an optionallysubstituted non-aromatic, three- to twelve-membered, cyclic hydrocarbonring, optionally containing one or more degrees of unsaturation, whichmay be optionally substituted as herein further described, with multipledegrees of substitution being allowed. Exemplary “cycloalkyl” groups asused herein include, but are not limited to, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, and cycloheptyl, as well as rings containingone or more degrees of unsaturation but short of aromatic, such ascyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, andcycloheptenyl.

As used herein, the term “cycloalkylene” refers to a divalent,non-aromatic cyclic hydrocarbon ring, which may be optionallysubstituted as herein further described, with multiple degrees ofsubstitution being allowed. Exemplary “cycloalkylene” groups include,but are not limited to, cyclopropylene, cyclobutylene, cyclopentylene,cyclohexylene, cycloheptylene, and substituted versions thereof. Theterm is intended to encompass divalent rings having different points ofattachment as well as a common point of attachment, which connectingatom may also be referred to as “spiroatom.”

As used herein, the term “heterocycle” or “heterocyclyl” refers to anoptionally substituted mono- or polycyclic ring system, optionallycontaining one or more degrees of unsaturation and also containing oneor more heteroatoms, which may be optionally substituted as hereinfurther described, with multiple degrees of substitution being allowed.Exemplary heteroatoms include nitrogen, oxygen, or sulfur atoms,including N-oxides, sulfur oxides, and sulfur dioxides. Typically, thering is three to twelve-membered and is either fully saturated or hasone or more degrees of unsaturation. Such rings may be optionally fusedto one or more of another heterocyclic ring(s), cycloalkyl ring(s), arylgroups (as defined below) or heteroaryl groups (as defined below).Examples of “heterocyclic” groups as used herein include, but are notlimited to, tetrahydrofuran, pyran, 1,4-dioxane, 1,3-dioxane,piperidine, pyrrolidine, morpholine, tetrahydrothiopyran, andtetrahydrothiophene.

As used herein, the term “aryl” refers to a univalent aromatic ring orfused ring system, which may be optionally substituted as herein furtherdescribed, with multiple degrees of substitution being allowed. Examplesof “aryl” groups as used include, but are not limited to, phenyl,2-naphthyl, 1-naphthyl, anthracene, and phenanthrene.

As used herein, the term “heteroaryl” refers to a monocyclic five toseven membered aromatic ring, or to a fused bicyclic aromatic ringsystem comprising two of such aromatic rings, which may be optionallysubstituted as herein further described, with multiple degrees ofsubstitution being allowed. These heteroaryl rings contain one or morenitrogen, sulfur, and/or oxygen atoms, where N-oxides, sulfur oxides,and dioxides are permissible heteroatom substitutions. Examples of“heteroaryl” groups as used herein include, but should not be limitedto, furan, thiophene, pyrrole, imidazole, pyrazole, triazole, tetrazole,thiazole, oxazole, isoxazole, oxadiazole, thiadiazole, isothiazole,pyridine, pyridazine, pyrazine, pyrimidine, quinoline, isoquinoline,benzofuran, benzodioxolyl, benzothiophene, indole, indazole,benzimidizolyl, imidazopyridinyl, pyrazolopyridinyl, andpyrazolopyrimidinyl.

As used herein, the term “fused cycloalkylaryl” refers to one or twocycloalkyl groups fused to an aryl group, the aryl and cycloalkyl groupshaving two atoms in common, and wherein the aryl group is the point ofsubstitution. Examples of “fused cycloalkylaryl” used herein include5-indanyl, 5,6,7,8-tetrahydro-2-naphthyl,

and the like.

As used herein, the term “fused arylcycloalkyl” refers to one or twoaryl groups fused to a cycloalkyl group, the cycloalkyl and aryl groupshaving two atoms in common, and wherein the cycloalkyl group is thepoint of substitution. Examples of “fused arylcycloalkyl” used hereininclude 1-indanyl, 2-indanyl, 9-fluorenyl,1-(1,2,3,4-tetrahydronaphthyl),

and the like.

As used herein, the term “fused heterocyclylaryl” refers to one or twoheterocyclyl groups fused to an aryl group, the aryl and heterocyclylgroups having two atoms in common, and wherein the aryl group is thepoint of substitution. Examples of “fused heterocyclylaryl” used hereininclude 3,4-methylenedioxy-1-phenyl,

and the like.

As used herein, the term “fused arylheterocyclyl” refers to one or twoaryl groups fused to a heterocyclyl group, the heterocyclyl and arylgroups having two atoms in common, and wherein the heterocyclyl group isthe point of substitution. Examples of “fused arylheterocyclyl” usedherein include 2-(1,3-benzodioxolyl),

and the like.

As used herein, the term “fused cycloalkylheteroaryl” refers to one ortwo cycloalkyl groups fused to a heteroaryl group, the heteroaryl andcycloalkyl groups having two atoms in common, and wherein the heteroarylgroup is the point of substitution. Examples of “fusedcycloalkylheteroaryl” used herein include 5-aza-6-indanyl,

and the like.

As used herein, the term “fused heteroarylcycloalkyl” refers to one ortwo heteroaryl groups fused to a cycloalkyl group, the cycloalkyl andheteroaryl groups having two atoms in common, and wherein the cycloalkylgroup is the point of substitution. Examples of “fusedheteroarylcycloalkyl” used herein include 5-aza-1-indanyl,

and the like.

As used herein, the term “fused heterocyclylheteroaryl” refers to one ortwo heterocyclyl groups fused to a heteroaryl group, the heteroaryl andheterocyclyl groups having two atoms in common, and wherein theheteroaryl group is the point of substitution. Examples of “fusedheterocyclylheteroaryl” used herein include1,2,3,4-tetrahydro-beta-carbolin-8-yl,

and the like.

As used herein, the term “fused heteroarylheterocyclyl” refers to one ortwo heteroaryl groups fused to a heterocyclyl group, the heterocyclyland heteroaryl groups having two atoms in common, and wherein theheterocyclyl group is the point of substitution. Examples of “fusedheteroarylheterocyclyl” used herein include-5-aza-2,3-dihydrobenzofuran-2-yl,

and the like.

As used herein the term “alkoxy” refers to a group —OR^(a), where R^(a)is alkyl as defined above.

As used herein the term “halogen” refers to fluorine, chlorine, bromine,or iodine.

As used herein the term “haloalkyl” refers to an alkyl group, as definedherein, that is substituted with at least one halogen. Examples ofbranched or straight chained “haloalkyl” groups as used herein include,but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, andt-butyl substituted independently with one or more halogens, forexample, fluoro, chloro, bromo, and iodo. The term “haloalkyl” should beinterpreted to include such substituents as perfluoroalkyl groups suchas —CF₃.

As used herein, the term “haloalkylene” refers to a straight or branchedchain divalent hydrocarbon radical, substituted with at least onehalogen. The temr should be interpreted to include perfluoroalkylenegroups such as —CF₂—.

As used herein, the term “haloalkoxy” refers to a group —OR^(a), whereR^(a) is a haloalkyl group as herein defined. As non-limiting examples,haloalkoxy groups include —O(CH₂)F, —O(CH)F₂, and —OCF₃.

As used herein the term “nitro” refers to a group —NO₂.

As used herein the term “cyano” refers to a group —CN.

As used herein the term “azido” refers to a group —N₃.

As used herein the term “amide” refers to a group —C(O)NR^(a)R^(b) or—NR^(a)C(O)—, where each R^(a) and R^(b) individually is hydrogen,alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocylcyl, or heteroaryl.

As used herein “amino” refers to a group —NR^(a)R^(b), where each ofR^(a) and R^(b) individually is hydrogen, alkyl, alkenyl, alkynyl,cycloalkyl, aryl, heterocylcyl, or heteroaryl. As used herein, wheneither R^(a) or R^(b) is other than hydrogen, such a group may bereferred to as a “substituted amino” or, for example if R^(a) is H andR^(b) is alkyl, as an “alkylamino.”

As used herein, the term “acyl” refers to a group —C(O)R^(a), whereR^(a) is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl,heterocyclyl, or heteroaryl.

As used herein, the term “hydroxyl” refers to a group —OH.

As used herein, the term “substituted” refers to substitution of one ormore hydrogens of the designated moiety with the named substituent orsubstituents, multiple degrees of substitution being allowed unlessotherwise stated, provided that the substitution results in a stable orchemically feasible compound. A stable compound or chemically feasiblecompound is one in which the chemical structure is not substantiallyaltered when kept at a temperature from about −80° C. to about +40° C.,in the absence of moisture or other chemically reactive conditions, forat least a week, or a compound which maintains its integrity long enoughto be useful for therapeutic or prophylactic administration to apatient. As used herein, the phrase “substituted with one or more . . .”refers to a number of substituents that equals from one to the maximumnumber of substituents possible based on the number of available bondingsites, provided that the above conditions of stability and chemicalfeasibility are met.

The present invention includes a compound of Formula (I):

or a salt thereof, wherein

-   A¹ is O or S;-   A² is O or S;-   U is N or C—(CH₂)_(u)R^(U);-   V is N or C—(CH₂)_(v)R^(V);-   W is N or C—(CH₂)_(w)R^(W);-   X is N or C—(CH₂)_(x)R^(X);-   Y is N or C—(CH₂)_(y)R^(Y);-   Z is N or C—(CH₂)_(z)R^(Z);-   R¹ is —(CH₂)_(q)R^(Q);-   R² is —(CH₂)_(s)R^(S);-   wherein    -   each of q, s, u, v, w, x, y, and z individually is 0, 1, 2, 3,        or 4;    -   each of R^(Q), R^(S), R^(U), R^(V), R^(W), R^(X), R^(Y), and        R^(Z) is independently selected from the group consisting of        -   alkyl;        -   alkyl substituted with one or more R^(ak);        -   alkenyl;        -   alkynyl;        -   aryl;        -   aryl substituted with one or more R^(a);        -   azido;        -   cyano;        -   cycloalkyl;        -   cycloalkyl substituted with one or more R^(c);        -   fused cycloalkylaryl substituted with one or more R^(f1);        -   fused arylcycloalkyl substituted with one or more R^(f2);        -   fused heterocyclyaryl substituted with one or more R^(f3);        -   fused arylheterocyclyl substituted with one or more R^(f4);        -   fused cycloalkylheteroaryl substituted with one or more            R^(f5);        -   fused heteroarylcycloalkyl substituted with one or more            R^(f6);        -   fused heterocyclylheteroaryl substituted with one or more            R^(f7);        -   fused heteroarylheterocyclyl substituted with one or more            R^(f8);        -   halogen;        -   haloalkyl;        -   heterocyclyl;        -   heterocyclyl substituted with one or more R6 ^(hc);        -   heteroaryl;        -   heteroaryl substituted with one or more R^(ha);        -   hydrogen;        -   —NR³R⁴;        -   —C(O)NR³R⁴;        -   —C(O)R⁵;        -   —C(O)₂R⁶;        -   —S(O)_(j)R⁷;        -   —OR⁸; and        -   nitro;        -   wherein            -   each of R³ and R⁴ is independently selected from the                group consisting of H, acyl, alkyl, alkoxy, alkoxyalkyl,                alkylsulfonyl, aryl, cycloalkyl, heterocyclyl and                heteroaryl;            -   j is 0, 1, or 2;            -   each R⁵, R⁶, R⁷, and R⁸ is independently selected from                the group consisting of                -   hydrogen;                -   alkyl;                -   alkyl substituted with one or more R^(ak);                -   alkenyl;                -   alkynyl;                -   alkoxy;                -   aryl;                -   aryl substituted with one or more R^(a);                -   cycloalkyl;                -   cycloalkyl substituted with one or more R^(c);                -   halogen;                -   haloalkyl;                -   heterocyclyl;                -   heterocyclyl substituted with one or more R^(hc);                -   heteroaryl;                -   heteroaryl substituted with one or more R^(ha);                -   wherein each R^(a), R^(c), R^(f1), R^(f2), R^(f3),                    R^(f4), R^(f5), R^(f6), R^(f7), R^(hc) and R^(ha) is                    independently selected from the group consisting of                    acyl, alkyl, alkenyl, alkynyl, alkoxy, amide, amino,                    aryl, cyano, cycloalkyl, halogen, haloalkyl,                    haloalkoxy, heteroaryl, hydroxy, nitro, —C(O)OR⁹,                    —SO₂R¹⁰, —SR¹¹, —C(O)R¹², —C(O)NR¹³, —NH—SO₂—R¹⁴,                    —SO₂—NR¹⁵R¹⁶, and —SO₂—CH₂—SO₂—CH₃; and wherein                -    each R⁹, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, R¹⁵, and R¹⁶ is                    independently selected from the group consisting of                    hydrogen, alkyl, alkenyl, alkynyl, aryl, cycloalkyl,                    heterocyclyl, and heteroaryl,-   provided that when R¹ is hydrogen, R^(S) is selected from the group    consisting of    -   alkyl;    -   alkyl substituted with one or more R^(ak);    -   alkenyl;    -   alkynyl;    -   aryl;    -   aryl substituted with one or more R^(a);    -   azido;    -   cyano;    -   cycloalkyl;    -   cycloalkyl substituted with one or more R^(c);    -   fused cycloalkylaryl substituted with one or more R^(f1);    -   fused arylcycloalkyl substituted with one or more R^(f2);    -   fused heterocyclyaryl substituted with one or more R^(f3);    -   fused arylheterocyclyl substituted with one or more R^(f4);    -   fused cycloalkylheteroaryl substituted with one or more R^(f5);    -   fused heteroarylcycloalkyl substituted with one or more R^(f6);    -   fused heterocyclylheteroaryl substituted with one or more        R^(f7);    -   fused heteroarylheterocyclyl substituted with one or more        R^(f8);    -   heterocyclyl;    -   heterocyclyl substituted with one or more R^(hc);    -   heteroaryl;    -   heteroaryl substituted with one or more R^(ha);    -   —NR³R⁴;    -   —C(O)NR³R⁴;    -   —C(O)R⁵;    -   —C(O)₂R⁶;    -   —S(O)_(j)R⁷;    -   —OR⁸; and    -   nitro.

In one embodiment, U is N.

In another embodiment, A¹ is O.

In another embodiment, A² is O.

In another embodiment, both A¹ and A² are O.

In another embodiment x is zero.

In another embodiment y is zero.

In another embodiment, V is C—(CH₂)_(v)R^(V); W is C—(CH₂)_(w)R^(W); Xis C—(CH₂)_(x)R^(X); Y is C—(CH₂)_(y)R^(Y); and Z is C—(CH₂)_(z)R^(Z).

In another embodiment, V is C—(CH₂)_(v)R^(V), R^(V) is halogen or —O—CH₃and v is zero.

In another embodiment, X is C—(CH₂)_(x)R^(X), R^(X) is halogen orhydrogen and x is zero.

In another embodiment, at least one of v, w and z is 1. In anotherembodiment, at least one of R^(W), R^(X) and R^(Y) is halogen. Inanother embodiment, w is zero and R^(W) is chloro or fluoro. In anotherembodiment, x is zero and R^(X) is chloro or fluoro.

In another embodiment, R¹ is —(CH₂)_(q)R^(Q), where q is 0 and R^(Q) isalkyl; aryl; aryl substituted with one or more alkoxy; cycloalkyl;cycloalkyl substituted with one or more alkyl, halogen, or —C(O)OR⁹,where R⁹ is hydrogen; alkyl; haloalkyl; heterocyclyl; or heterocyclylsubstituted with one or more acyl, —C(O)OR⁹, or —S(O)₂R¹⁰, where each R⁹and R¹⁰ independently is alkyl or aryl. In one embodiment, q is 1, R^(Q)is aryl, aryl substituted with alkoxy, or heterocyclyl. In oneembodiment, q is 3, R^(Q) is heterocyclyl substituted with —OR⁸ and R⁸is alkyl.

In another embodiment, R² is —(CH₂)_(s)R^(S), where s is 0 and R^(S) isalkyl; aryl; aryl substituted with one or more alkyl, alkoxy, halogen,haloalkyl, or nitro; cycloalkyl; heteroaryl; or heteroaryl substitutedwith one or more alkyl or haloalkyl. In one embodiment, s is 1 and R^(S)is aryl, aryl substituted with one or more alkyl, or heteroaryl. In oneembodiment, s is 1 or 2 and R^(S) is —C(O)OR⁶, where R⁶ is alkyl. In oneembodiment, s is 2 and R^(S) is aryl.

In another embodiment, U is —C—OR⁸. In another embodiment, R⁸ is alkyl.In yet another embodiment, R⁸ is methyl. In one embodiment, R^(U) is H,alkyl, or haloalkoxy.

In another embodiment, R^(S) is aryl. In another embodiment, R^(S) is asubstituted or unsubstituted phenyl group. In still another embodiment,R^(S) is a chloro-substituted phenyl group.

In another embodiment, R^(S) is a benzodioxoyl group. In anotherembodiment, R^(S) is a benzo[1,3]dioxolyl group. In one embodiment,R^(S) is a benzodioxinyl group. In another embodiment, R^(S) is abenzo[1,4]dioxinyl group.

In another embodiment, R^(Q) is a substituent having the structure ofFormula (R^(Q))

wherein M is nitrogen or carbon and T is nitrogen, carbon, oxygen,sulfur or SO₂, p is from 1 to 3 and t is from 1 to 3, wherein R_(Q) maybe substituted or unsubstituted as described above. In anotherembodiment, R_(Q) is a piperdinyl group, where M is nitrogen, T is—N(H)—, and t and p are both 2.

The present invention also includes a compound of Formula (Ia):

or a salt thereof, wherein

-   A¹ is O or S;-   A² is O or S;-   R¹ is hydrogen or —(CH₂)_(q)R^(Q);-   R² is hydrogen or —(CH₂)_(s)R^(S);-   R¹⁷ is hydrogen or —(CH₂)_(v)R^(V);-   R¹⁸ is hydrogen or —(CH₂)_(w)R^(W);-   R¹⁹ is hydrogen or —(CH₂)_(x)R^(X);-   R²⁰ is hydrogen or —(CH₂)_(y)R^(Y);-   wherein    -   each of q, s, v, w, x, and y individually is 0, 1, 2, 3, or 4;    -   each of R^(Q), R^(S), R^(V), R^(W), R^(X), and R^(Y) is        independently selected from the group consisting of        -   alkyl;        -   alkyl substituted with one or more R^(ak);        -   alkenyl;        -   alkynyl;        -   aryl;        -   aryl substituted with one or more R^(a);        -   azido;        -   cyano;        -   cycloalkyl;        -   cycloalkyl substituted with one or more R^(c);        -   fused cycloalkyaryl;        -   fused cycloalkylaryl substituted with one or more R^(f1);        -   fused arylcycloalkyl;        -   fused arylcycloalkyl substituted with one or more R^(f2);        -   fused heterocyclylaryl;        -   fused heterocyclyaryl substituted with one or more R^(f3);        -   fused arylheterocycyl;        -   fused arylheterocyclyl substituted with one or more R^(f4);        -   fused cycloalkylheteroaryl;        -   fused cycloalkylheteroaryl substituted with one or more            R^(f5);        -   fused heteroarylcycloalkyl;        -   fused heteroarylcycloalkyl substituted with one or more            R^(f6);        -   fused heterocyclylheteroaryl;        -   fused heterocyclylheteroaryl substituted with one or more            R^(f7);        -   fused heteroarylheterocyclyl;        -   fused heteroarylheterocyclyl substituted with one or more            R^(f8);        -   halogen;        -   haloalkyl;        -   heterocyclyl;        -   heterocyclyl substituted with one or more R^(hc);        -   heteroaryl;        -   heteroaryl substituted with one or more R^(ha);        -   —NR³R⁴;        -   —C(O)NR³R⁴;        -   —C(O)R⁵;        -   —C(O)₂R⁶;        -   —S(O)_(j)R⁷;        -   —OR⁸; and        -   nitro;        -   wherein            -   each of R³ and R⁴ is independently selected from the                group consisting of H, acyl, alkyl, alkoxy, alkoxyalkyl,                alkylsulfonyl, aryl, cycloalkyl, heterocyclyl and                heteroaryl;            -   j is 0, 1, or 2;            -   each R⁵, R⁶, R⁷, and R⁸ is independently selected from                the group consisting of                -   hydrogen;                -   alkyl;                -   alkyl substituted with one or more R^(ak);                -   alkenyl;                -   alkynyl;                -   alkoxy;                -   aryl;                -   aryl substituted with one or more R^(a);                -   cycloalkyl;                -   cycloalkyl substituted with one or more R^(c);                -   halogen;                -   haloalkyl;                -   heterocyclyl;                -   heterocyclyl substituted with one or more R^(hc);                -   heteroaryl;                -   heteroaryl substituted with one or more R^(ha);                -   wherein each R^(a), R^(c), R^(f1), R^(f2), R^(f3),                    R^(f4), R^(f5), R^(f6), R^(f7), R^(hc) and R^(ha) is                    independently selected from the group consisting of                    acyl, alkyl, alkenyl, alkynyl, alkoxy, amide, amino,                    aryl, cyano, cycloalkyl, halogen, haloalkyl,                    haloalkoxy, heteroaryl, hydroxy, nitro, —C(O)OR⁹,                    —SO₂R¹⁰, —SR¹¹, —C(O)R¹², —C(O)NR¹³, —NH—SO₂—R¹⁴,                    —SO₂—NR¹⁵R¹⁶, and —SO₂—CH₂—SO₂—CH₃; and wherein                -    each R⁹, R¹⁰, R¹¹, R¹², R¹³ R¹⁴, R¹⁵, and R¹⁶ is                    independently selected from the group consisting of                    hydrogen, alkyl, alkenyl, alkynyl, aryl, cycloalkyl,                    heterocyclyl, and heteroaryl,-   provided that when R¹ is hydrogen, R^(S) is selected from the group    consisting of    -   alkyl;    -   alkyl substituted with one or more R^(ak);    -   alkenyl;    -   alkynyl;    -   aryl;    -   aryl substituted with one or more R^(a);    -   azido;    -   cyano;    -   cycloalkyl;    -   cycloalkyl substituted with one or more R^(c);    -   fused cycloalkylaryl substituted with one or more R^(f1);    -   fused arylcycloalkyl substituted with one or more R^(f2);    -   fused heterocyclyaryl substituted with one or more R^(f3);    -   fused arylheterocyclyl substituted with one or more R^(f4);    -   fused cycloalkylheteroaryl substituted with one or more R^(f5);    -   fused heteroarylcycloalkyl substituted with one or more R^(f6);    -   fused heterocyclylheteroaryl substituted with one or more        R^(f7);    -   fused heteroarylheterocyclyl substituted with one or more        R^(f8);    -   heterocyclyl;    -   heterocyclyl substituted with one or more R^(hc);    -   heteroaryl;    -   heteroaryl substituted with one or more R^(ha);    -   —NR³R⁴;    -   —C(O)NR³R⁴;    -   —C(O)R⁵;    -   —C(O)₂R⁶;    -   —S(O)_(j)R⁷;    -   —OR⁸; and    -   nitro.

The present invention also includes a compound of Formula (Ib):

or a salt thereof, wherein R¹, R², R^(V), R^(W), R^(X), R^(Y), v, w, xand y are as defined with regard to Formula (I).

In one embodiment of Formula (Ib), R^(Y), R^(X) and R^(W) are hydrogenor halogen and y, x and w are each zero.

In another embodiment of Formula (Ib), R^(Y), R^(X) and R^(W) arehydrogen or halogen, y, x and w are each zero, v is zero, and R^(V) isalkyl, haloalkyl, cycloalkyl, halogen or —OR¹², where R¹² is hydrogen,alkyl or haloalkyl.

In another embodiment of Formula (Ib), R¹ is hydrogen, alkyl,cycloalkyl, phenyl, benzyl, tetrahydrofuranyl, tetrahydropyranyl,pyrrolidinyl or piperidinyl, wherein, the alkyl, cycloalkyl,piperidinyl, pyrrolindinyl, phenyl and benzyl may be each independentlysubstituted with one or more R^(a), as described with reference toFormula (I).

In another embodiment of Formula (Ib), R² is hydrogen alkyl, cycloalkyl,phenyl, benzyl, furanyl, thiophenyl, isoxazolyl, thiophenyl, furanyl,benzodioxolyl, dihydrobenzodioxinyl, indanyl, wherein, the alkyl,cycloalkyl, phenyl, benzyl, thiophenyl and furanyl may be eachindependently substituted with one or more R^(a), as described withreference to Formula (I).

In another embodiment of Formula (Ib), R^(Y), R^(X) and R^(W) arehydrogen or halogen, y, x and w are each zero, v is zero, and R^(V) isalkyl, haloalkyl, halogen or —OR¹², where R¹² is hydrogen, alkyl orhaloalkyl, and R¹ is —(CH₂)_(q)—R^(Q), wherein q is zero and R^(Q)selected from the group consisting of: hydrogen; C₃-C₆ cycloalkyl; C₃-C₆cycloalkyl substituted with one or more C₁-C₆ alkyl, halogen, —C(O)OR⁹wherein R⁹ is hydrogen or C₁-C₃ alkyl, or —NH—SO₂—R¹⁴ wherein R¹⁴ isC₁-C₃ alkyl; C₁-C₆ alkyl; C₁-C₆ alkyl substituted with one or morehalogen, C₁-C₆ alkoxy, C₁-C₆ haloalkyl, pyrrolidin-1-yl, or —SO₂—R¹⁰wherein R¹⁰ is C₁-C₃ alkyl; phenyl; phenyl substituted with one or moreone or more halogen, C₁-C₆ alkyl, or C₁-C₆ alkoxy; —C(O)—R⁵, wherein R⁵is phenyl; —C(O)—R⁵, wherein R⁵ is phenyl substituted with one or morehalogen, C₁-C₆ alkyl, or C₁-C₆ alkoxy; tetrahydro-furan-3-yl;piperidine-4-yl; piperidine-4-yl substituted at the 1 position with—C(O)O—R⁹ wherein R⁹ is C₁-C₆ alkyl, —C(O)—R¹² wherein R¹² is C₁-C₆alkyl or phenyl, —SO₂—R¹⁰ wherein R¹⁰ is C₁-C₆ alkyl, phenyl,1,1-dioxo-tetrahydro-1-lambda-6-thiophene or —CH₂—SO₂—CH₃, —SO₂—NR¹⁵R¹⁶are independently selected from the group consisting of hydrogen andC₁-C₆ alkyl, or —C(O)NR¹³ wherein R¹³ is C₁-C₆ alkyl; pyrrolidine-3-yl;and pyrrolidine-3-yl substituted at the 1 position with —C(O)O—R⁹wherein R⁹ is C₁-C₆ alkyl, or —SO₂—R¹⁰ wherein R¹⁰ is C₁-C₆ alkyl orphenyl; or R¹ is —(CH₂)_(q)—R^(Q), wherein q is 1 and R^(Q) selectedfrom the group consisting of: phenyl; phenyl substituted with one ormore halogen, C₁-C₆ alkyl, or C₁-C₆ alkoxy; tetrahydro-furan-2-yl;piperidine-4-yl; piperidine-4-yl substituted at the 1 position with—SO₂—R¹⁰ wherein R¹⁰ is C₁-C₆ alkyl; and tetrahydro-pyran-4-yl; and R²is —(CH₂)_(s)—R^(S), wherein s is zero and R^(S) selected from the groupconsisting of: hydrogen; C₃-C₆ cycloalkyl; C₁-C₆ alkyl; C₁-C₆ alkylsubstituted with phenyl or —C(O)OR⁹ wherein R⁹ is C₁-C₃ alkyl; phenyl;phenyl substituted with one or more halogen, C₁-C₆ alkyl, C₁-C₆haloalkyl, C₁-C₆ alkoxy; C₁-C₆ haloalkoxy, nitrile, —S—R¹¹ wherein R¹¹is C₁-C₆ alkyl, or —SO₂—R¹⁰ wherein R¹⁰ is C₁-C₆ alkyl; furan-3-yl;furan-3-yl substituted with one or more C₁-C₆ alkyl, or C₁-C₆ haloalkyl;thiophen-2-yl; thiophen-2-yl substituted with one or more C₁-C₆ alkyl;thiophen-3-yl; 3,5-dimethyl-isoxazol-4-yl; 3-benzo[1,3]dioxol-5-yl;3-indan-5-yl; and 2,3-dihydro-benzo [1,4]dioxin-6-yl; or R² is—(CH₂)_(s)—R^(S), wherein s is 1 and R^(S) selected from the groupconsisting of phenyl; phenyl substituted with one or more halogen, C₁-C₆alkyl, C₁-C₆ alkoxy, or C₁-C₆ haloalkoxy; —C(O)OR⁹, wherein R⁹ is C₁-C₃alkyl; and furan-2-yl. In a further embodiment, at least one of R¹ or R²is not hydrogen. In a further embodiment, neither R¹ not R² is hydrogen.

In another embodiment of Formula (Ib), R^(Y), R^(X) and R^(W) arehydrogen or halogen, y, x and w are each zero, v is zero, and R^(V) isalkyl, haloalkyl, halogen or —OR¹², where R¹² is hydrogen, alkyl orhaloalkyl, and R¹ is —(CH₂)_(q)—R^(Q), wherein q is zero and R^(Q)selected from the group consisting of: cyclohexyl; cyclohexylsubstituted with one or more C₁-C₆ alkyl, halogen, —C(O)OR⁹ wherein R⁹is hydrogen or C₁-C₃ alkyl, or —NH—SO₂—R¹⁴ wherein R¹⁴ is C₁-C₃ alkyl;C₁-C₆ alkyl; C₁-C₆ alkyl substituted with one or more halogen, —SO₂—R¹⁰wherein R¹⁰ is C₁-C₃ alkyl; phenyl; phenyl substituted with one or moreone or more halogen, C₁-C₆ alkyl, or C₁-C₆ alkoxy; —C(O)—R⁵, wherein R⁵is phenyl; —C(O)—R⁵, wherein R⁵ is phenyl substituted with one or morehalogen, C₁-C₆ alkyl, or C₁-C₆ alkoxy; piperidine-4-yl; piperidine-4-ylsubstituted at the 1 position with —C(O)O—R⁹ wherein R⁹ is C₁-C₆ alkyl,—C(O)—R¹² wherein R¹² is C₁-C₆ alkyl, —SO₂—R¹⁰ wherein R¹⁰ is C₁-C₆alkyl or phenyl, —SO₂—NR¹⁵R¹⁶ wherein R¹⁵ and R¹⁶ are independentlyselected from the group consisting of hydrogen and C₁-C₆ alkyl, or—C(O)NR¹³ wherein R¹³ is C₁-C₆ alkyl; or R¹ is —(CH₂)_(q)—R^(Q), whereinq is 1 and R^(Q) selected from the group consisting of: phenyl; phenylsubstituted with one or more halogen, C₁-C₆ alkyl, or C₁-C₆ alkoxy;piperidine-4-yl; piperidine-4-yl substituted at the 1 position with—SO₂—R¹⁰ wherein R¹⁰ is C₁-C₆ alkyl; and R² is —(CH₂)_(s)—R^(S), whereins is zero and R^(S) selected from the group consisting of: C₃-C₆cycloalkyl; C₁-C₆ alkyl; phenyl; phenyl substituted with one or morehalogen, C₁-C₆ alkyl, C₁-C₆ alkoxy; C₁-C₆ haloalkoxy, nitrile, —S—R¹¹wherein R¹¹ is C₁-C₆ alkyl, or —SO₂—R¹⁰ wherein R¹⁰ is C₁-C₆ alkyl; orR² is —(CH₂)_(s)—R^(S), wherein s is 1 and R^(S) selected from the groupconsisting of phenyl; and phenyl substituted with one or more halogen,C₁-C₆ alkyl, C₁-C₆ alkoxy; or C₁-C₆ haloalkoxy.

In another embodiment of Formula (Ib), R^(Y), R^(X) and R^(W) arehydrogen or halogen, y, x and w are each zero, v is zero, and R^(V) isalkyl, haloalkyl, halogen or —OR¹², where R¹² is hydrogen, alkyl orhaloalkyl, and R¹ is —(CH₂)_(q)—R^(Q), wherein q is zero and R^(Q)selected from the group consisting of: cyclohexyl; cyclohexylsubstituted with one or more C₁-C₆ alkyl, halogen, —C(O)OR⁹ wherein R⁹is hydrogen or C₁-C₃ alkyl; C₁-C₆ alkyl; C₁-C₆ alkyl substituted withone or more halogen; phenyl; phenyl substituted with one or more one ormore halogen, C₁-C₆ alkyl, or C₁-C₆ alkoxy; and R² is —(CH₂)_(s)—R^(S),wherein s is zero and R^(S) selected from the group consisting of: C₁-C₆alkyl; phenyl; phenyl substituted with one or more halogen, C₁-C₆ alkyl,C₁-C₆ alkoxy; C₁-C₆ haloalkoxy; or R² is —(CH₂)_(s)—R^(S), wherein s is1 and R^(S) selected from the group consisting of phenyl; and phenylsubstituted with one or more halogen, C₁-C₆ alkyl, C₁-C₆ alkoxy; orC₁-C₆ haloalkoxy.

The present invention also includes a compound of Formula (Ic):

or a salt thereof, wherein A¹, A², R², R^(V), R^(W), R^(X), R^(Y), v, w,x and y are as defined with regard to Formula (I).

The present invention also includes a compound of Formula (Id):

or a salt thereof, wherein A¹, A², R¹, R^(V), R^(W), R^(X), R^(Y), v, w,x and y are as defined with regard to Formula (I).

Another embodiment of the present invention includes a compound ashereinbefore described with reference to any one of the below-identifiedExamples, or a salt thereof.

Another embodiment of the present invention includes a pharmaceuticalcomposition comprising a compound of the present invention. Anotherembodiment of the present invention includes a pharmaceuticalcomposition further including one or more pharmaceutically acceptablecarriers. Another embodiment of the present invention includes apharmaceutical composition further comprising one or more additionaltherapeutic agent, yet another embodiment includes wherein the one ormore additional therapeutic agent is selected from steroids,cyclooxygenase inhibitors, non-steroidal-anti-inflammatory drugs, orTNF-α antibodies, such as for example acetyl salicylic acid, bufexamac,diclofenac potassium, sulindac, diclofenac sodium, ketorolac trometamol,tolmetine, ibuprofen, naproxen, naproxen sodium, tiaprofen acid,flurbiprofen, mefenamic acid, nifluminic acid, meclofenamate,indomethacin, proglumetacine, ketoprofen, nabumetone, paracetamol,piroxicam, tenoxicam, nimesulide, fenylbutazon, tramadol, beclomethasonedipropionate, betamethasone, beclamethasone, budesonide, fluticasone,mometasone, dexamethasone, hydrocortisone, methylprednisolone,prednisolone, prednisone, triamcinolone, celecoxib, rofecoxib,infliximab, leflunomide, etanercept, CPH 82, methotrexate,sulfasalazine, antilymphocytory immunoglobulines, antithymocytoryimmunoglobulines, azathioprine, cyclosporine, tacrolimus substances,ascomycin, rapamycin, or muromonab-CD3.

Another embodiment of the present invention includes a compound of thepresent invention for use as an active therapeutic substance.

Another embodiment of the present invention includes a compound of thepresent invention for use to inhibit the activity of TNF-α in a subjectin need thereof.

Another embodiment of the present invention includes a compound of thepresent invention for use to inhibit PDE4 in a subject in need thereof.

Another embodiment of the present invention includes a compound of thepresent invention for use in the treatment or prevention of conditionsor disorders mediated by activity of TNF-α.

Another embodiment of the present invention includes a compound of thepresent invention for use in the treatment or prevention of conditionsor disorders mediated by PDE4.

Another embodiment of the present invention includes a use of a compoundof the present invention in the manufacture of a medicament for use ofinhibiting the activity of TNF-α in a subject in need thereof.

Another embodiment of the present invention includes a use of a compoundof the present invention in the manufacture of a medicament for use ofinhibiting PDE4 in a subject in need thereof.

Another embodiment of the present invention includes a use of a compoundof the present invention in the manufacture of a medicament for use inthe treatment or prevention of conditions or disorders mediated byactivity of TNF-α.

Another embodiment of the present invention includes a use of a compoundof the present invention in the manufacture of a medicament for use inthe treatment or prevention of conditions or disorders mediated by PDE4.

Another embodiment of the present invention includes a method ofinhibiting the activity of TNF-α in a subject in need thereof throughthe administration of a compound of the present invention.

Another embodiment of the present invention includes a method ofinhibiting PDE4 in a subject in need thereof through the administrationof a compound of the present invention.

Another embodiment of the present invention includes a method for thetreatment or prevention of conditions or disorders mediated by activityof TNF-α through the administration of a compound of the presentinvention.

Another embodiment of the present invention includes a method for thetreatment or prevention of conditions or disorders mediated by PDE4through the administration of a compound of the present invention.

Another embodiment of the present invention includes a method for thetreatment of an inflammatory disease through the administration of acompound of the present invention. Inflammatory diseases may include, asnon-limiting examples, viral, alcoholic, or drug-induced acute andfulminant hepatitis, hepatic steatosis, both alcoholic andnon-alcoholic, viral and non-viral hepatitis, hepatic cirrhosis,autoimmune hepatitis, chronic active hepatitis, Wilson's disease,myasthenia gravis, idiopathic sprue, autoimmune inflammatory boweldisease, ulcerative colitis, Crohn's disease, inflammatory boweldiseases, endocrine opthalmopathy, Grave's disease, sarcoidosis, primarybiliary cirrhosis, pancreatitis, nephritis, endotoxin shock, septicshock, haemodynamic shock, sepsis syndrome, post ischemic reperfusioninjury, malaria, mycobacterial infection, meningitis, psoriasis, asthma,chronic obstructive pulmonary disease (COPD), eosinophilia, congestiveheart failure, fibrotic diseases, cystic fibrosis, pulmonary fibrosis,hepatic fibrosis, renal fibrosis, cachexia, graft rejection, graft vs.host disease, rejection by transplantation, cancer, diseases involvingangiogenesis, autoimmune diseases, ankylosing spondylitis, autoimmuneencephalomyelitis, autoimmune hematological disorders, hemolytic anemia,aplastic anemia, pure red cell anemia, idiopathic thrombocytopenia,systemic lupus erythematosus (SLE), polychondritis, scleroderma, Wegenergranulomatosis, dermatomyositis, Reiter's syndrome, non infectionuveitis, autoimmune keratitis, keratoconjunctivitis sicca, vernalkeratoconjunctivitis, interstitial lung fibrosis, psoriatic arthritis,psoriasis and other benign or malignant proliferative skin diseases,atopic dermatitis, urticaria, neurodegenerative disorders, Parkinson'sdisease, Alzheimer's disease, acute and chronic multiple sclerosis,cancer, viral infection, human immunodeficiency virus (HIV), cachexia,thrombosis, skin inflammatory diseases, osteoarthritis (OA),osteoporosis, RA, emphysema, chronic bronchiolitis, allergic rhinitis,radiation damage, hyperoxic alveolar injury, periodontal disease,non-insulin dependent diabetes mellitus (Type II diabetes), and insulindependent diabetes mellitus (Juvenile or Type I diabetes).

In another embodiment, such treatment or prevention relates toconditions mediated by the inhibition of PDE 4. Such conditions includea variety of conditions, especially inflammatory and/or allergicdiseases, in mammals such as humans, for example: asthma, chronicobstructive pulmonary disease (COPD) (e.g. chronic bronchitis and/oremphysema), atopic dermatitis, urticaria, allergic rhinitis, allergicconjunctivitis, vernal conjunctivitis, eosinophilic granuloma,psoriasis, rheumatoid arthritis, septic shock, ulcerative colitis,Crohn's disease, reperfusion injury of the myocardium and brain, chronicglomerulonephritis, endotoxic shock, adult respiratory distresssyndrome, multiple sclerosis, cognitive impairment (e.g. in aneurological disorder such as Alzheimer's disease), depression, or pain.Ulcerative colitis and/or Crohn's disease are collectively oftenreferred to as inflammatory bowel disease.

In on embodiment of the present invention, the inflammatory and/orallergic disease is chronic obstructive pulmonary disease (COPD),asthma, rheumatoid arthritis or allergic rhinitis in a mammal (e.g.human).

The scope of the present invention includes combinations of embodiments.

Unless otherwise stated, structures depicted herein are also meant toinclude compounds which differ only in the presence of one or moreisotopically enriched atoms. For example, compounds having the presentstructure except for the replacement of a hydrogen atom by a deuteriumor tritium, or the replacement of a carbon atom by a ¹³C- or¹⁴C-enriched carbon are within the scope of the invention.

Certain of the compounds described herein contain one or more chiralcenters, or may otherwise be capable of existing as multiplestereoisomers. The scope of the present invention includes mixtures ofstereoisomers as well as purified enantiomers orenantiomerically/diastereomerically enriched mixtures. Also includedwithin the scope of the invention are the individual isomers of thecompounds represented by the formulae of the present invention, as wellas any wholly or partially equilibrated mixtures thereof. The presentinvention also includes the individual isomers of the compoundsrepresented by the formulas above as mixtures with isomers thereof inwhich one or more chiral centers are inverted.

The present invention includes salts of the compounds herein described.

The term “salt” or “pharmaceutically acceptable salt” refers to saltsthat retain the biological effectiveness and properties of the compoundsof Formula I, and which are not biologically or otherwise undesirable.Pharmaceutically acceptable base addition salts can be prepared frominorganic and organic bases. Salts derived from inorganic bases, includeby way of example only, sodium, potassium, lithium, ammonium, calciumand magnesium salts. Salts derived from organic bases include, but arenot limited to, salts of primary, secondary and tertiary amines, such asalkyl amines, dialkyl amines, trialkyl amines, substituted alkyl amines,di(substituted alkyl) amines, tri(substituted alkyl) amines, alkenylamines, dialkenyl amines, trialkenyl amines, substituted alkenyl amines,di(substituted alkenyl) amines, tri(substituted alkenyl) amines,cycloalkyl amines, di(cycloalkyl) amines, tri(cycloalkyl) amines,substituted cycloalkyl amines, disubstituted cycloalkyl amine,trisubstituted cycloalkyl amines, cycloalkenyl amines, di(cycloalkenyl)amines, tri(cycloalkenyl) amines, substituted cycloalkenyl amines,disubstituted cycloalkenyl amine, trisubstituted cycloalkenyl amines,aryl amines, diaryl amines, triaryl amines, heteroaryl amines,diheteroaryl amines, triheteroaryl amines, heterocyclic amines,diheterocyclic amines, triheterocyclic amines, mixed di- and tri-amineswhere at least two of the substituents on the amine are different andare selected from the group consisting of alkyl, substituted alkyl,alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl, heterocyclic,and the like. Also included are amines where the two or threesubstituents, together with the amino nitrogen, form a heterocyclic orheteroaryl group.

Specific examples of suitable amines include, by way of example only,isopropylamine, trimethyl amine, diethyl amine, tri(iso-propyl) amine,tri(n-propyl) amine, ethanolamine, 2-dimethylaminoethanol, tromethamine,lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline,betaine, ethylenediamine, glucosamine, N-alkylglucamines, theobromine,purines, piperazine, piperidine, morpholine, N-ethylpiperidine, and thelike.

Salts of the compounds of the present invention may further compriseacid addition salts. Representative salts include acetate,benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate,calcium edetate, camsylate, carbonate, clavulanate, citrate,dihydrochloride, edisylate, estolate, esylate, fumarate, gluceptate,gluconate, glutamate, glycollylarsanilate, hexylresorcinate,hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide,isethionate, lactate, lactobionate, laurate, malate, maleate, mandelate,mesylate, methylsulfate, monopotassium maleate, mucate, napsylate,nitrate, N-methylglucamine, oxalate, pamoate (embonate), palmitate,pantothenate, phosphate/diphosphate, polygalacturonate, potassium,salicylate, sodium, stearate, subacetate, succinate, sulfate, tannate,tartrate, teoclate, tosylate, triethiodide, trimethylammonium, andvalerate salts.

As used herein, the phrase “effective amount” means that amount of adrug or pharmaceutical agent that will elicit the biological or medicalresponse of a tissue, system, animal, or human that is being sought, forinstance, by a researcher or clinician. The phrase “therapeuticallyeffective amount” means any amount which, as compared to a correspondingsubject who has not received such amount, results in improved treatment,healing, prevention, or amelioration of a disease, disorder, or sideeffect, or a decrease in the rate of advancement of a disease ordisorder. The term also includes within its scope amounts effective toenhance normal physiological function. For use in therapy,therapeutically effective amounts of a compound of formulae of thepresent, as well as salts thereof, may be administered as the rawchemical. Additionally, the active ingredient may be presented as apharmaceutical composition.

A therapeutically effective amount of a compound of the presentinvention will depend upon a number of factors. For example, thespecies, age and weight of the recipient, the precise conditionrequiring treatment and its severity, the nature of the formulation andthe route of administration are all factors to be considered. Thetherapeutically effective amount ultimately should be at the discretionof the attendant or veterinarian. Regardless, an effective amount of acompound of the present invention for the treatment of humans,generally, should be in the range of 0.01 to 100 mg/kg body weight ofrecipient (mammal) per day. More usually the effective amount should bein the range of 1 to 10 mg/kg body weight per day. Thus, for a 70 kgadult mammal the actual amount per day would usually be from 70 to 700mg. This amount may be given in a single dose per day or in a number(such as two, three, four, five, or more) of sub-doses per day such thatthe total daily dose is the same. An effective amount of a salt may bedetermined as a proportion of the effective amount of the compound ofthe formulae of the present invention per se. Similar dosages should beappropriate for treatment of the other conditions referred to herein.

The compounds of the present invention demonstrate utility as inhibitorsof TNF-α activity. In an embodiment, the invention provides a method forinhibiting TNF-α activity comprising contacting a cell in whichinhibition of TNF-α is desired with a TNF-α inhibitor of the presentinvention. In an embodiment, the TNF-α inhibitor interacts with andreduces the activity of TNF-α in a cell.

As used herein, reference is made to one or more disorder, condition, ordisease mediated by TNF-α or amplified by TNF-α and such shall includediseases associated with or implicating TNF-α activity, for example, theoveractivity of TNF-α, and conditions that accompany with these diseaseswith reference to: 1) TNF-α expression in cells which normally do notexpress TNF-α; 2) TNF-α activity by cells which normally do not possessactive TNF-α; 3) increased TNF-α expression leading to unwanted cellproliferation; or 4) mutations leading to constitutive activation ofTNF-α.

The present invention further provides pharmaceutical compositions thatinclude effective amounts of compounds of the formulae of the presentinvention and salts thereof, and one or more pharmaceutically acceptablecarriers, diluents, or excipients. The compounds of the formulae of thepresent invention, including salts thereof, are as herein described. Thecarrier(s), diluent(s), or excipient(s) must be acceptable, in the senseof being compatible with the other ingredients of the formulation andnot deleterious to the recipient of the pharmaceutical composition.

In accordance with another aspect of the invention there is alsoprovided a process for the preparation of a pharmaceutical formulationincluding admixing a compound of the formulae of the present invention,including a salt thereof, with one or more pharmaceutically acceptablecarriers, diluents or excipients.

Pharmaceutical formulations may be presented in unit dose formscontaining a predetermined amount of active ingredient per unit dose.Such a unit may contain, as a non-limiting example, 0.5 mg to 1 g of acompound of the present invention, depending on the condition beingtreated, the route of administration, and the age, weight, and conditionof the patient. Typical unit dosage formulations are those containing adaily dose or sub-dose, as herein above recited, or an appropriatefraction thereof, of an active ingredient. Such pharmaceuticalformulations may be prepared by any of the methods well known in thepharmacy art.

Pharmaceutical formulations may be adapted for administration by anyappropriate route, for example by an oral (including buccal orsublingual), rectal, nasal, topical (including buccal, sublingual ortransdermal), vaginal, or parenteral (including subcutaneous,intramuscular, intravenous or intradermal) route. Such formulations maybe prepared by any method known in the art of pharmacy, for example bybringing into association the active ingredient with the carrier(s) orexcipient(s). By way of example, and not meant to limit the invention,with regard to certain conditions and disorders for which the compoundsof the present invention are believed useful, certain routes will bepreferable to others.

Pharmaceutical formulations adapted for oral administration may bepresented as discrete units such as capsules or tablets; powders orgranules; solutions or suspensions, each with aqueous or non-aqueousliquids; edible foams or whips; or oil-in-water liquid emulsions orwater-in-oil liquid emulsions. For instance, for oral administration inthe form of a tablet or capsule, the active drug component can becombined with an oral, non-toxic pharmaceutically acceptable inertcarrier such as ethanol, glycerol, water, and the like. Generally,powders are prepared by comminuting the compound to a suitable fine sizeand mixing with an appropriate pharmaceutical carrier such as an ediblecarbohydrate, as, for example, starch or mannitol. Flavorings,preservatives, dispersing agents, and coloring agents can also bepresent.

Capsules are made by preparing a powder, liquid, or suspension mixtureand encapsulating with gelatin or some other appropriate shell material.Glidants and lubricants such as colloidal silica, talc, magnesiumstearate, calcium stearate, or solid polyethylene glycol can be added tothe mixture before the encapsulation. A disintegrating or solubilizingagent such as agar-agar, calcium carbonate or sodium carbonate can alsobe added to improve the availability of the medicament when the capsuleis ingested. Moreover, when desired or necessary, suitable binders,lubricants, disintegrating agents, and coloring agents can also beincorporated into the mixture. Examples of suitable binders includestarch, gelatin, natural sugars such as glucose or beta-lactose, cornsweeteners, natural and synthetic gums such as acacia, tragacanth, orsodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, andthe like.

Lubricants useful in these dosage forms include, for example, sodiumoleate, sodium stearate, magnesium stearate, sodium benzoate, sodiumacetate, sodium chloride, and the like. Disintegrators include, withoutlimitation, starch, methyl cellulose, agar, bentonite, xanthan gum, andthe like.

Tablets are formulated, for example, by preparing a powder mixture,granulating or slugging, adding a lubricant and disintegrant, andpressing into tablets. A powder mixture may be prepared by mixing thecompound, suitably comminuted, with a diluent or base as describedabove. Optional ingredients include binders such ascarboxymethylcellulose, aliginates, gelatins, or polyvinyl pyrrolidone,solution retardants such as paraffin, resorption accelerators such as aquaternary salt, and/or absorption agents such as bentonite, kaolin, ordicalcium phosphate. The powder mixture can be wet-granulated with abinder such as syrup, starch paste, acadia mucilage or solutions ofcellulosic or polymeric materials, and forcing through a screen. As analternative to granulating, the powder mixture can be run through thetablet machine and the result is imperfectly formed slugs broken intogranules. The granules can be lubricated to prevent sticking to thetablet-forming dies by means of the addition of stearic acid, a stearatesalt, talc or mineral oil. The lubricated mixture is then compressedinto tablets. The compounds of the present invention can also becombined with a free flowing inert carrier and compressed into tabletsdirectly without going through the granulating or slugging steps. Aclear or opaque protective coating consisting of a sealing coat ofshellac, a coating of sugar or polymeric material, and a polish coatingof wax can be provided. Dyestuffs can be added to these coatings todistinguish different unit dosages.

Where appropriate, dosage unit formulations for oral administration canbe microencapsulated. The formulation can also be prepared to prolong orsustain the release as for example by coating or embedding particulatematerial in polymers, wax or the like.

Oral fluids such as solutions, syrups, and elixirs can be prepared indosage unit form so that a given quantity contains a predeterminedamount of the compound. Syrups can be prepared, for example, bydissolving the compound in a suitably flavored aqueous solution, whileelixirs are prepared through the use of a non-toxic alcoholic vehicle.Suspensions can be formulated generally by dispersing the compound in anontoxic vehicle. Solubilizers and emulsifiers such as ethoxylatedisostearyl alcohols and polyoxy ethylene sorbitol ethers, preservatives;flavor additives such as peppermint oil, or natural sweeteners,saccharin, or other artificial sweeteners; and the like can also beadded.

Suitable packaging for the pharmaceutical solution formulations may beall approved containers intended for parenteral use, such as plastic andglass containers, ready-to-use syringes and the like. In an embodiment,the container is a sealed glass container, such as a vial or an ampoule.A hermetically sealed glass vial is one example of a sealed glasscontainer. According to an embodiment of the present invention, there isprovided, in a sealed glass container, a sterile, injectable solutioncomprising a compound of the present invention in a physiologicallyacceptable solvent, and which has an appropriate pH for stability. Acidsalts of the compounds of the present invention may be more soluble inaqueous solutions than their free base counter parts, but when the acidsalts are added to aqueous solutions the pH of the solution may be toolow to be suitable for administration. Thus, solution formulationshaving a pH above pH 4.5 may be combined prior to administration with adiluent solution of pH greater than 7 such that the pH of thecombination formulation administered is pH 4.5 or higher. In oneembodiment, the diluent solution comprises a pharmaceutically acceptablebase such as sodium hydroxide and the pH of the combined formulationadministered is between pH 5.0 and 7.0. One or more additionalcomponents such as co-solubilizing agents, tonicity adjustment agents,stabilizing agents and preservatives, for instance of the kindpreviously specified, may be added to the solution prior to passing thesolution through the sterilizing filter.

Pharmaceutical formulations adapted for topical administration may beformulated as ointments, creams, suspensions, lotions, powders,solutions, pastes, gels, sprays, aerosols, or oils.

For treatments of the eye or other external tissues, for example mouthand skin, the formulations may be applied as a topical ointment orcream. When formulated in an ointment, the active ingredient may beemployed with either a paraffinic or a water-miscible ointment base.Alternatively, the active ingredient may be formulated in a cream withan oil-in-water cream base or a water-in-oil base. Pharmaceuticalformulations adapted for topical administrations to the eye include eyedrops wherein the active ingredient is dissolved or suspended in asuitable carrier, especially an aqueous solvent.

Pharmaceutical formulations adapted for topical administration in themouth include lozenges, pastilles, and mouthwashes.

Pharmaceutical formulations adapted for nasal administration, where thecarrier is a solid, include a coarse powder having a particle size forexample in the range 20 to 500 microns. The powder is administered inthe manner in which snuff is taken, i.e., by rapid inhalation throughthe nasal passage from a container of the powder held close up to thenose. Suitable formulations wherein the carrier is a liquid, foradministration as a nasal spray or as nasal drops, include aqueous oroil solutions of the active ingredient.

Pharmaceutical formulations adapted for administration by inhalationinclude fine particle dusts or mists, which may be generated by means ofvarious types of metered dose pressurized aerosols, nebulizers, orinsufflators.

Further, the compositions of the present invention may also be in theform of suppositories for rectal administration of the compounds of theinvention. These compositions can be prepared by mixing the drug with asuitable non-irritating excipient which is solid at ordinarytemperatures but liquid at the rectal temperature and will thus melt inthe rectum to release the drug. Such materials include cocoa butter andpolyethylene glycols, for example. Pharmaceutical formulations adaptedfor rectal administration may be presented as suppositories or asenemas.

Pharmaceutical formulations adapted for vaginal administration may bepresented as pessaries, tampons, creams, gels, pastes, foams, or sprayformulations.

Pharmaceutical formulations adapted for parenteral administrationinclude aqueous and non-aqueous sterile injection solutions which maycontain anti-oxidants, buffers, bacteriostats, and solutes that renderthe formulation isotonic with the blood of the intended recipient; andaqueous and non-aqueous sterile suspensions which may include suspendingagents and thickening agents. The formulations may be presented inunit-dose or multi-dose containers, for example sealed ampules andvials, and may be stored in a freeze-dried (lyophilized) conditionrequiring only the addition of the sterile liquid carrier, for examplewater for injections, immediately prior to use.

Extemporaneous injection solutions and suspensions may be prepared fromsterile powders, granules, and tablets.

In addition to the ingredients particularly mentioned above, theformulations may include other agents conventional in the art havingregard to the type of formulation in question. For example, formulationssuitable for oral administration may include flavoring or coloringagents.

In a further embodiment to this aspect and described hereinbelow infurther detail, the invention encompasses a combination therapy fortreating or preventing a disorder mediated by TNF-α in a subject. Thecombination therapy comprises administering to the subject atherapeutically or prophylactically effective amount of a compound ofthe present invention and one or more other therapy includingchemotherapy, radiation therapy, gene therapy, and immunotherapy.

In an embodiment of the present invention, the compound of the presentinvention may be administered in combination with other therapeuticcompounds. In particular, a compound of this invention can beadvantageously used in combination with i) Leukotriene receptorantagonists, ii) Leukotriene biosynthesis inhibitors, iii) COX-2selective inhibitors, iv) statins, v) NSAIDs, vi) M2/M3 antagonists,vii) corticosteroids, viii) Hi (histamine) receptor antagonists, ix)beta 2 adrenoceptor agonists, x) interferons, xi) antiviral drugs suchas protease inhibitors, helicase inhibitors, polymerase inhibitors,lamivudine, and the like, xiii) ursodesoxycholic acid, xiv)glycyrrhizin, xv) human growth factor (HGF), or xvi) aminosalicylicacids such as salazosulfapyridine, mesalazin, or the like.

The compounds of the present invention may also be used in combinationwith other conventional anti-inflammatory or immunosuppressive agents,such as steroids, cyclooxygenase inhibitors,non-steroidal-anti-inflammatory drugs, TNF-α antibodies or otherTNF-binding proteins, such as for example acetyl salicylic acid,bufexamac, diclofenac potassium, sulindac, diclofenac sodium, ketorolactrometamol, tolmetine, ibuprofen, naproxen, naproxen sodium, tiaprofenacid, flurbiprofen, mefenamic acid, nifluminic acid, meclofenamate,indomethacin, proglumetacine, ketoprofen, nabumetone, paracetamol,piroxicam, tenoxicam, nimesulide, fenylbutazon, tramadol, beclomethasonedipropionate, betamethasone, beclamethasone, budesonide, fluticasone,mometasone, dexamethasone, hydrocortisone, methylprednisolone,prednisolone, prednisone, triamcinolone, celecoxib, rofecoxib,infliximab, leflunomide, etanercept, CPH 82, methotrexate,sulfasalazine, antilymphocyte immunoglobulins, antithymocyteimmunoglobulins, azathioprine, cyclosporine, tacrolimus substances,ascomycin, rapamycin, adalimumab, muromonab-CD3 or other antibodies orfusion proteins that modulate T-cell function such as abatacept,alefacept and efalizumab. Additionally, yet to emerge agents arecontemplated as being useful in combination with the compound of thepresent invention.

As noted above, the compounds of the present invention may be employedalone or in combination with other therapeutic agents, including othercompounds of the present invention. Such a combination ofpharmaceutically active agents may be administered together orseparately and, when administered separately, administration may occursimultaneously or sequentially, in any order. The amounts of thecompounds or agents and the relative timings of administration will beselected in order to achieve the desired therapeutic effect. Theadministration in combination of a compound of the formulae of thepresent invention including salts thereof with other treatment agentsmay be in combination by administration concomitantly in: (1) a unitarypharmaceutical composition including both compounds; or (2) separatepharmaceutical compositions each including one of the compounds.Alternatively, the combination may be administered separately in asequential manner wherein one treatment agent is administered first andthe other second or vice versa. Such sequential administration may beclose in time or remote in time. The compounds of the present inventionmay be used in the treatment of a variety of disorders and conditionsand, as such, the compounds of the present invention may be used incombination with a variety of other suitable therapeutic agents usefulin the treatment or prophylaxis of those disorders or conditions.

The compounds of this invention may be made by a variety of methods,including well-known standard synthetic methods. An illustrative generalsynthetic method is set out in the following reaction Scheme (in whichvariables are as defined before or are defined) using readily availablestarting materials, and reagents. In these reactions, it is alsopossible to make use of variants which are themselves known to those ofordinary skill in this art, but are not mentioned in greater detail.Specific compounds of the invention are prepared in the workingExamples.

In all of the examples described below, protecting groups for sensitiveor reactive groups are employed where necessary in accordance withgeneral principles of synthetic chemistry. Protecting groups may bemanipulated according to standard methods of organic synthesis. Thesegroups may be removed at a convenient stage of the compound synthesisusing methods that are readily apparent to those skilled in the art. Theselection of processes as well as the reaction conditions and order oftheir execution shall be consistent with the preparation of compounds ofthe present invention.

Those skilled in the art will recognize if a stereocenter exists. Asnoted hereinabove, the present invention includes all possiblestereoisomers and includes not only racemic compounds but the individualenantiomers as well. When a compound is desired as a single enantiomer,such may be obtained by stereospecific synthesis, by resolution of thefinal product or any convenient intermediate, or by chiralchromatographic methods as are known in the art. Resolution of the finalproduct, an intermediate, or a starting material may be affected by anysuitable method known in the art.

The present invention also provides a method for the synthesis ofcompounds useful as intermediates in the preparation of compounds of thepresent invention along with methods for their preparation.

The compounds can be prepared according to the following Generalprocedures. In these reactions, variants may be employed which arethemselves known to those of ordinary skill in this art, but are notmentioned in greater detail. As will be apparent to those skilled in theart, the format and designation of variable substituent groups in thefollowing procedures may be different from that used in the Formulae ofthe present invention.

Description of General Synthetic Methods

General procedure A: Preparation of 4-chloro-quinoline-3-carboxylic acidethyl ester analogues

To a substituted aniline (5 mmol) in chlorobenzene (15 mL) is addeddiethyl ethoxymethylene-malonate (5 mmol) and stirred at 140° C. for12-16 h. The reaction mass is concentrated to dryness on a rotaryevaporator, POCl₃ (15 mL) is added and the reaction mass is stirred at120° C. for 12 h or microwaved at 150° C. for 30 min. After removingmost of POCl₃ on rotary evaporator, a slurry of ice in sodiumbicarbonate solution is slowly added and the resulting reaction productis extracted into ethyl acetate (2×50 mL). The combined ethyl acetateextracts are washed with brine, dried over sodium sulfate, concentratedand purified on column to give the desired quinoline derivative.

The following 4-chloro-quinoline-3-carboxylic acid ethyl ester analogueswere prepared using the above procedure:

-   4,6-Dichloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester;-   4,7-Dichloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester;-   4-Chloro-6-fluoro-8-methoxy-quinoline-3-carboxylic acid ethyl ester;-   4-Chloro-8-ethyl-quinoline-3-carboxylic acid ethyl ester;-   4-Chloro-8-propyl-quinoline-3-carboxylic acid ethyl ester;-   4-Chloro-8-isopropyl-quinoline-3-carboxylic acid ethyl ester; and-   4-Chloro-8-trifluoromethoxy-quinoline-3-carboxylic acid ethyl ester.    General procedure B: Preparation of 4-alkylamino- or    4-arylamino-quinoline-3-carboxylic acid ethyl ester analogues

To a solution of a 4-chloro-quinoline-3-carboxylic acid ethyl esteranalogue (10.0 mmol) in anhydrous THF (50 mL) is added cesium carbonate(20.0 mmol) and an amine (1.2 eq. 12.0 mmol). The reaction mixture isthen heated to reflux and stirred at 70° C. for 4 h. After completion ofthe reaction, the mixture is poured into water (100 mL) and extractedwith ethyl acetate (2×50 mL) and the organic extracts are combined,washed with brine solution (2×50 mL), concentrated in vacuo to give thecrude product. The concentrated residue is then purified with silica gelchromatography using hexanes-ethyl acetate to afford4-amino-quinoline-3-carboxylic acid ethyl ester derivative.

General procedure C: Preparation of substituted1H-pyrimido[5,4-c]quinoline-2,4-diones and substituted1H-pyrimido[5,4-c]quinoline-thione-ketone derivatives

To a solution of a 4-amino-quinoline-3-carboxylic acid ethyl esterderivative (0.50 mmol) in 10 mL anhydrous DMF is added sodium hydride (4eq., 2.0 mmol) and stirred at room temperature for 30 min. To thissolution an isocyanate or isothiocyanate (1.5-4 eq. 2.0 mmol) is added.The resulting reaction mixture is stirred for 18 h at room temperatureor at 100° C. After completion of the reaction, the mixture is pouredinto water (20 mL) and extracted with ethyl acetate (2×25 mL). Theorganic extracts are combined, washed with water (2×25 mL), brine (2×25mL), dried over Na₂SO₄ and concentrated in vacuo to give the crudeproduct. The residue is then purified by silica gel chromatography usingdichloromethane-ethyl acetate to provide desired1H-pyrimido[5,4-c]quinoline-2,4-dione or thio-ketone derivative.

General procedure D: Ester hydrolysis

4-Alkylamino or 4-arylamino-quinoline-3-carboxylic acid ethyl ester(0.50 mmol) is dissolved in THF/MeOH (3 mL, 2:1) and 10 N aq NaOHsolution (0.15 mL, 1.5 mmol) is added. After stirring for 1 h at 60° C.,the reaction mass is concentrated to dryness, acidified with 4N HCl indioxane, and concentrated to dryness. To this solid is added 10%methanol in dichloromethane and sodium sulfate. The solution is stirredat room temperature (r.t.), filtered through filter paper and washedwith 10% methanol in dichloromethane. The combined filtrates areconcentrated to dryness to give acid.

General procedure E: Preparation of amides

4-Alkylamino or 4-arylamino-quinoline-3-carboxylic acid analogue (0.5mmol) and diisopropylethylamine (2.5 mmol) are dissolved in DMF (1 mL)and HBTU (0.6 mmol) is added to the solution. After 15-30 min ofstirring, an amine (0.65 mmol) is added and stirred for 30 min. Thereaction mixture is diluted with water and the product is extracted intoethyl acetate (2×10 mL). The combined ethyl acetate extracts are washedwith brine, dried over sodium sulfate and concentrated to give theamide.

General procedure F: Preparation of substituted1H-pyrimido[5,4-c]quinoline-2,4-diones

To an amide (0.5 mmol) in dry THF (or in dry DMF) is added NaH (2.0mmol) and stirred for 30 min at 60° C. Methyl chloroformate (1.0 mmol)is slowly added and continued stirring at 60° C. for 24 h. The reactionmass is cooled to room temperature, quenched excess NaH with water,diluted with water and extracted into ethyl acetate (2×20 mL). Thecombined ethyl acetate extracts are washed with brine, dried over sodiumsulfate, concentrated and purified on a silica gel column to givedesired substituted 1H-pyrimido[5,4-c]quinoline-2,4-dione.

General procedure G: Removal of tert-butyl carbamate

To a stirred solution of carbamate (1 mmol) in DCM-MeOH (4:1, 1 mL) isadded 4 N HCl in dioxane (5 mL). The reaction is stirred at roomtemperature for 30 min. Solvents are removed under reduced pressure andthe residue is dried under vacuum the product.

General procedure H: Preparation of amides, sulfonamides, sulfamides,ureas and carbamates

To a stirred solution of amine (1 mmol) in DCM (1 mL) at 0° C. is addedtriethylamine (3 mmol) followed by addition of an acid chloride orsulfonyl chloride or sulfamyl chloride or chloroformate or isocyanate(1.2 to 1.5 mmol) under N₂ atmosphere. The resultant reaction is stirredfor 2 h at room temperature. The reaction mixture is diluted with DCM(10 mL), washed with water (2×5 mL), dried over Na₂SO₄ and concentratedunder reduced pressure. The crude product is purified by silica gelflash column chromatography.

General procedure I: Acid-catalyzed hydrolysis of esters

To an ester (1 mmol) is added 4N HCl in dioxane (2 mL) and water (0.5mL) and stirred at 60° C. for 12 h. The reaction mixture is concentratedand azeotroped with toluene three times. The obtained crude is filteredon a small silica gel bed to give the desired acid.

General procedure J: Oxidation of sulfides to sulfones

To solution of a sulfide (0.1 mmol) in dichloromethane (DCM) (5 mL) isadded peracetic acid (15 μL, 0.2 mmol, 2.0 eq) at 0° C. and the reactionmixture is slowly warmed up to room temperature while stirring for 30min. Upon completion of the reaction, the mixture is poured intosaturated aqueous sodium bicarbonate (10 mL) solution, extracted withDCM (2×25 mL). The organic extracts are combined and washed with brine(1×25 mL) and concentrated under reduced pressure. The residue is thenpurified with silica gel chromatography using DCM:EtOAc to give thedesired sulfone.

General procedure K: Cyclization

To the amino-ester (1 mmol) in DMSO (5 mL) was added isocyanate (2 mmol)and the contents were heated at 100° C. for 4 h. Cooled to roomtemperature and added methanol (10 mL) and concentated. To the residuewas added DBU (1 mmol) and heated for 10 min at 60° C. Cooled thereaction mass to room temperature, diluted with water and extracted intoethyl acetate (2×20 mL). Combined ethyl acetate extracts were washedwith brine, dried over sodium sulfate, concentrated and purified onsilicagel column to give desired substituted1H-pyrimido[5,4-c]quinoline-2,4-dione.

EXAMPLES Example 11-cyclopentyl-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (2.65 g, 10.0mmol) was treated with cyclopentylamine(1.06 g, 1.2 eq. 12.0 mmol)following a method similar to general procedure B to afford4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(2.9 g). The obtained amino-ester (0.5 mmol) was subjected to reactionwith ethyl isocyanate (2.0 mmol) according to general procedure C tofurnish1-cyclopentyl-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(83 mg). LCMS: m/z 340 [M+1]⁺. ¹H NMR (400 MHz, CDCl₃): δ

9.43 (s, 1H), 7.78 (d, 1H), 7.55 (t, 1H), 7.20 (d, 1H), 4.99 (p, 1H),4.13 (s, 3H), 4.11 (q, 2H), 2.42 (m, 2H), 2.06 (m, 4H), 1.62 (m, 2H) and1.32 (t, 3H) ppm.

Example 21-Cyclopentyl-3-ethyl-7-methoxy-2-thioxo-2,3-dihydro-1H-pyrimido[5,4-c]quinolin-4-one

1-Cyclopentyl-3-ethyl-7-methoxy-2-thioxo-2,3-dihydro-1H-pyrimido[5,4-c]quinolin-4-one(12 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.10 mmol) (prepared as described in Example 1) and ethylisothiocyanate (0.4 mmol) following a procedure similar to generalprocedure C. LCMS: m/z 356 [M+1]⁺. ¹H NMR (400 MHz, CDCl₃): δ 9.50 (s,1H), 7.74 (d, 1H), 7.53 (t, 1H), 7.21 (d, 1H), 5.03 (p, 1H), 4.09 (s,3H), 4.07 (q, 2H), 2.42 (m, 2H), 2.08 (m, 4H), 1.64 (m, 2H) and 1.34 (t,3H).

Example 31,3-Dicyclopentyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1,3-Dicyclopentyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione (17mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.10 mmol) and cyclopentyl isocyanate (0.4 mmol) following generalprocedure C. LCMS: m/z 380 [M+1]⁺. ¹H NMR (400 MHz, CDCl₃): δ 9.51 (s,1H), 7.82 (d, 1H), 7.62 (t, 1H), 7.28 (d, 1H), 5.34 (m, 1H), 4.98 (m,1H), 4.15 (s, 3H), 3.95 (m, 4H), 2.38 (m, 2H), 2.12 (m, 2H), 1.93-1.98(m, 2H), 1.58-169 (m, 4H), and 1.38 (m, 2H) ppm.

Example 41-(3,4-Dimethoxy-phenyl)-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (89 mg, 0.3mmol) was treated with 3,4-dimethoxy-aniline following general procedureB to afford4-(3,4-dimethoxy-phenylamino)-8-methoxy-quinoline-3-carboxylic acidethyl ester (103 mg). Thus obtained amino-ester (38 mg, 0.1 mmol) wassubjected to reaction with ethyl isocyanate (0.5 mmol) according togeneral procedure C to furnish1-(3,4-dimethoxy-phenyl)-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(23 mg). LCMS: m/z 408 [M+1]⁺. ¹H NMR (400 MHz, CDCl₃): δ 9.64 (s, 1H),7.11 (m, 2H), 7.00 (s, 1H), 6.91 (m, 2H), 6.60 (s, 1H), 4.20 (q, 2H),4.09 (s, 3H), 3.99 (s, 3H), 3.86 (s, 3H), and 1.37 (t, 3H) ppm.

Example 51-Cyclopentyl-3-isopropyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-3-isopropyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(19 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.10 mmol) and isopropyl isocyanate (0.4 mmol) following generalprocedure C. LCMS: m/z 354 [M+1]⁺. ¹H NMR (400 MHz, CDCl₃): δ 9.45 (s,1H), 7.78 (d, 1H), 7.57 (t, 1H), 7.24 (d, 1H), 5.21 (p, 1H), 4.93 (m,1H), 4.14 (s, 3H), 3.83 (m, 2H), 2.38 (m, 2H), 2.08 (m, 4H), and 1.15(d, 6H) ppm.

Example 61-Cyclopentyl-7-methoxy-3-propyl-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-propyl-1H-pyrimido[5,4-c]quinoline-2,4-dione(14 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.10 mmol) and propyl isocyanate (0.4 mmol) following general procedureC. LCMS: m/z 354 [M+1]⁺. ¹H NMR (400 MHz, CDCl₃): δ 9.49 (s, 1H), 7.82(d, 1H), 7.59 (t, 1H), 7.24 (d, 1H), 5.01 (m, 1H), 4.15 (s, 3H), 4.03(t, 2H), 2.40 (m, 2H), 2.09 (m, 4H), 1.76 (m, 2H), 1.63 (m, 2H), and1.16 (t, 3H) ppm.

Example 73-Butyl-1-cyclopentyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-Butyl-1-cyclopentyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(26 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.10 mmol) and butyl isocyanate (0.4 mmol) following general procedureC. LCMS: m/z 368 [M+1]⁺. ¹H NMR (400 MHz, CDCl₃): δ 9.47 (s, 1H), 7.81(d, 1H), 7.57 (t, 1H), 7.23 (d, 1H), 4.99 (p, 1H), 4.14 (s, 3H), 4.06(t, 2H), 2.42 (m, 2H), 2.08 (m, 4H), 1.69 (m, 4H), 1.44 (q, 2H), 0.97(t, 3H) ppm.

Example 81-Cyclopentyl-3-furan-2-ylmethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-3-furan-2-ylmethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(21 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.10 mmol) and (2-furyl) methyl isocyanate (0.4 mmol) following generalprocedure C. LCMS: m/z 392 [M+1]⁺. ¹H NMR (400 MHz, CDCl₃): δ 9.45 (s,1H), 7.78 (d, 1H), 7.57 (t, 1H), 7.32 (d, 1H), 7.24 (d, 1H), 6.30 (d,1H), 6.18 (d, 1H), 5.04 (p, 1H), 4.33 (s, 2H), 4.12 (s, 3H), 2.40 (m,2H), 2.11 (m, 4H), and 1.16 (m, 2H) ppm.

Example 93-tert-Butyl-1-cyclopentyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-tert-Butyl-1-cyclopentyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(18 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.10 mmol) and tert-butyl isocyanate (0.4 mmol) following generalprocedure C. LCMS: m/z 368 [M+1]⁺.

Example 101-Cyclopentyl-7-methoxy-3-thiophen-2-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-thiophen-2-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione(19 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.10 mmol) and 2-isocyanatothiophene (0.4 mmol) following generalprocedure C. LCMS: m/z 394 [M+1]⁺.

Example 113-(1-Cyclopentyl-7-methoxy-2,4-dioxo-1,4-dihydro-2H-pyrimido[5,4-c]quinolin-3-yl)-propionicacid ethyl ester

4-(1-Cyclopentyl-7-methoxy-2,4-dioxo-1,4-dihydro-2H-pyrimido[5,4-c]quinolin-3-yl)-propionicacid ethyl ester (24 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.10 mmol) and 3-isocyanato-propionic acid ethyl ester (0.4 mmol)following general procedure C. LCMS: m/z 412 [M+1]⁺.

Example 121-Cyclopentyl-3-(2,4-dimethoxy-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-3-(2,4-dimethoxy-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(25 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.10 mmol) and 2,4-dimethoxyphenyl isocyanate (0.4 mmol) followinggeneral procedure C. LCMS: m/z 448 [M+1]⁺.

Example 13 1-Cyclopentyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione (8 mg) wasprepared from3-tert-butyl-1-cyclopentyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(18 mg, 0.05 mmol) with treatment of trifluoroacetic acid (20

L) in DCM (1 mL). LCMS: m/z 312 [M+1]⁺.

Example 141-Isopropyl-7-methoxy-3-propyl-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Isopropyl-7-methoxy-3-propyl-1H-pyrimido[5,4-c]quinoline-2,4-dione (23mg) was prepared from 4-isopropylamino-8-methoxy-quinoline-3-carboxylicacid ethyl ester (60 mg, 0.20 mmol) and n-propyl isocyanate (1.0 mmol)following general procedure C. LCMS: m/z 328 [M+1]⁺.

Example 151,3-Diisopropyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1,3-Diisopropyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione (21 mg)was prepared from 4-isopropylamino-8-methoxy-quinoline-3-carboxylic acidethyl ester (59.6 mg, 0.20 mmol) and isopropyl isocyanate (1.0 mmol)following general procedure C. LCMS: m/z 328 [M+1]⁺.

Example 163-Ethyl-7-methoxy-1-propyl-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg, 0.94mmol) was treated with 1-aminopropane following general procedure B toafford 8-methoxy-4-propylamino-quinoline-3-carboxylic acid ethyl ester(260 mg). Thus obtained amino-ester was hydrolyzed to the correspondingacid in quantitative yield using general procedure D and thentransformed into 8-methoxy-4-propylamino-quinoline-3-carboxylic acidethylamide (200 mg) following general procedure E. The above ethylamide(0.70 mmol) was subjected to reaction with methyl chloroformateaccording to general procedure F to furnish3-ethyl-7-methoxy-1-propyl-1H-pyrimido[5,4-c]quinoline-2,4-dione (26mg). LCMS: m/z 314 [M+1]⁺.

Example 171-Butyl-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (300 mg, 1.13mmol) was treated with 1-aminobutane following general procedure B toafford 4-butylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(295 mg). Thus obtained amino-ester was hydrolyzed to the correspondingacid in quantitative yield using general procedure D and thentransformed into the corresponding ethylamide (220 mg) following generalprocedure E.

The above ethylamide (0.73 mmol) was subjected to reaction with methylchloroformate according to general procedure F to furnish1-butyl-3-ethyl-7-methoxy-1H-pyrimido [5,4-c]quinoline-2,4-dione (70mg). LCMS: m/z 328 [M+1]⁺.

Example 183-Ethyl-1-isobutyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (300 mg, 1.13mmol) was treated with isobutylamine following general procedure B toafford 4-isobutylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(280 mg). Thus obtained amino-ester was hydrolyzed to the correspondingacid in quantitative yield using general procedure D and thentransformed into the corresponding ethylamide (180 mg) following generalprocedure E. The above ethylamide (0.6 mmol) was subjected to reactionwith methyl chloroformate according to general procedure F to furnish3-ethyl-1-isobutyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione (22mg). LCMS: m/z 328 [M+1]⁺.

Example 193-Ethyl-7-methoxy-1-(tetrahydro-furan-2-ylmethyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (300 mg, 1.13mmol) was treated with furfurylamine following general procedure B toafford8-methoxy-4-[(tetrahydro-furan-2-ylmethyl)-amino]-quinoline-3-carboxylicacid ethyl ester (280 mg). Thus obtained amino-ester was hydrolyzed tothe corresponding acid in quantitative yield using general procedure Dand then transformed into the corresponding ethylamide (220 mg)following general procedure E. The above ethylamide (0.67 mmol) wassubjected to reaction with methyl chloroformate according to generalprocedure F to furnish3-ethyl-7-methoxy-1-(tetrahydro-furan-2-ylmethyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(30 mg). LCMS: m/z 356 [M+1]⁺.

Example 20 1,3-Diethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (300 mg, 1.13mmol) was treated with ethylamine following general procedure B toafford 4-ethylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(220 mg). Thus obtained amino-ester was hydrolyzed to the correspondingacid in quantitative yield using general procedure D and thentransformed into the corresponding ethylamide (180 mg) following generalprocedure E. The above ethylamide (0.65 mmol) was subjected to reactionwith methyl chloroformate according to general procedure F to furnish1,3-diethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione (20 mg).LCMS: m/z 300 [M+1]⁺. LCMS: m/z 300 [M+1]⁺. ¹H NMR (400 MHz, CDCl₃): δ9.48 (s, 1H), 7.87 (d, 1H), 7.56 (t, 1H), 7.22 (d, 1H), 4.42 (q, 2H),4.16 (q, 2H), 4.12 (s, 3H), 1.73 (t, 3H), and 1.33 (t, 3H) ppm.

Example 211-Cyclopropyl-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (300 mg, 1.13mmol) was treated with cyclopropylamine following general procedure B toafford 4-cyclopropylamino-8-methoxy-quinoline-3-carboxylic acid ethylester (200 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid in quantitative yield using general procedure D andthen transformed into the corresponding ethylamide (170 mg) followinggeneral procedure E. The above ethylamide (0.6 mmol) was subjected toreaction with methyl chloroformate according to general procedure F tofurnish1-cyclopropyl-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(15 mg). LCMS: m/z 312 [M+1]⁺.

Example 221-Cyclobutyl-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (300 mg, 1.13mmol) was treated with cyclobutylamine following general procedure B toafford 4-cyclobutylamino-8-methoxy-quinoline-3-carboxylic acid ethylester (240 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid using general procedure D and then transformed intothe corresponding ethylamide (170 mg) following general procedure E.

The above ethylamide (0.60 mmol) was subjected to reaction with methylchloroformate according to general procedure F to furnish1-cyclobutyl-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione (20mg). LCMS: m/z 312 [M+1]⁺.

Example 233-Ethyl-1-(1-ethyl-propyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg, 0.94mmol) was treated with 1-ethyl-propylamine following general procedure Bto afford 4-(1-ethyl-propylamino)-8-methoxy-quinoline-3-carboxylic acidethyl ester (240 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid in quantitative yield using general procedure D andthen transformed into the corresponding ethylamide (160 mg) followinggeneral procedure E.

The above ethylamide (0.51 mmol) was subjected to reaction with methylchloroformate according to general procedure F to furnish3-ethyl-1-(1-ethyl-propyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(15 mg). LCMS: m/z 342 [M+1]⁺.

Example 243-Ethyl-7-methoxy-1-(tetrahydro-furan-3-yl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (300 mg, 1.13mmol) was treated with tetrahydro-furan-3-ylamine following generalprocedure B to afford8-methoxy-4-(tetrahydro-furan-3-ylamino)-quinoline-3-carboxylic acidethyl ester (275 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid in quantitative yield using general procedure D andthen transformed into the corresponding ethylamide (190 mg) followinggeneral procedure E. The above ethylamide (0.6 mmol) was subjected toreaction with methyl chloroformate according to general procedure F tofurnish3-ethyl-7-methoxy-1-(tetrahydro-furan-3-yl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(10 mg). LCMS: m/z 342 [M+1]⁺.

Example 253-Ethyl-7-methoxy-1-(3-methoxy-propyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg, 0.94mmol) was treated with 3-methoxy-propylamine following general procedureB to afford 8-methoxy-4-(3-methoxy-propyl-amino)-quinoline-3-carboxylicacid ethyl ester (260 mg). Thus obtained amino-ester was hydrolyzed tothe corresponding acid in quantitative yield using general procedure Dand then transformed into the corresponding ethylamide (180 mg)following general procedure E. The above ethylamide (0.57 mmol) wassubjected to reaction with methyl chloroformate according to generalprocedure F to furnish3-ethyl-7-methoxy-1-(3-methoxy-propyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(35 mg). LCMS: m/z 344 [M+1]⁺.

Example 261-(2,2-Dimethyl-propyl)-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg, 0.94mmol) was treated with 2,2-dimethyl-propylamine following generalprocedure B to afford4-(2,2-dimethyl-propylamino)-8-methoxy-quinoline-3-carboxylic acid ethylester (220 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid in quantitative yield using general procedure D andthen transformed into the corresponding ethylamide (160 mg) followinggeneral procedure E. The above ethylamide (0.50 mmol) was subjected toreaction with methyl chloroformate according to general procedure F tofurnish1-(2,2-dimethyl-propyl)-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(5 mg). LCMS: m/z 342 [M+1]⁺.

Example 273-Ethyl-7-methoxy-1-(4,4,4-trifluoro-butyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg, 0.94mmol) was treated with 4,4,4-trifluoro-butylamine following generalprocedure B to afford8-methoxy-4-(4,4,4-trifluoro-butylamino)-quinoline-3-carboxylic acidethyl ester (210 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid in quantitative yield using general procedure D andthen transformed into the corresponding ethylamide (170 mg) followinggeneral procedure E. The above ethylamide (0.48 mmol) was subjected toreaction with methyl chloroformate according to general procedure F tofurnish3-ethyl-7-methoxy-1-(4,4,4-trifluoro-butyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(70 mg). LCMS: m/z 382 [M+1]⁺.

Example 289-Chloro-1-cyclopentyl-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4,6-Dichloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg,0.83 mmol) was treated with cyclopentylamine following general procedureB to afford 6-chloro-4-cyclopentylamino-8-methoxy-quinoline-3-carboxylicacid ethyl ester (240 mg). Thus obtained amino-ester was hydrolyzed tothe corresponding acid in quantitative yield using general procedure Dand then transformed into the corresponding ethylamide (170 mg)following general procedure E.

The above ethylamide (0.49 mmol) was subjected to reaction with methylchloroformate according to general procedure F to furnish9-chloro-1-cyclopentyl-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(15 mg). LCMS: m/z 374 [M+1]⁺.

Example 291-Cyclopentyl-3-ethyl-9-fluoro-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-6-fluoro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250mg, 0.89 mmol) was treated with cyclopentylamine following generalprocedure B to afford4-cyclopentylamino-6-fluoro-8-methoxy-quinoline-3-carboxylic acid ethylester (200 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid in quantitative yield using general procedure D andthen transformed into the corresponding ethylamide (140 mg) followinggeneral procedure E. The above ethylamide (0.42 mmol) was subjected toreaction with methyl chloroformate according to general procedure F tofurnish1-cyclopentyl-3-ethyl-9-fluoro-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(13 mg). LCMS: m/z 358 [M+1]⁺.

Example 307-Chloro-1-cyclopentyl-3-ethyl-1H-pyrimido[5,4-c]quinoline-2,4-dione

4,8-Dichloro-quinoline-3-carboxylic acid ethyl ester (250 mg, 0.93 mmol)was treated with cyclopentylamine following general procedure B toafford 8-chloro-4-cyclopentylamino-quinoline-3-carboxylic acid ethylester (220 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid in quantitative yield using general procedure D andthen transformed into the corresponding ethylamide (165 mg) followinggeneral procedure E.

The above ethylamide (0.52 mmol) was subjected to reaction with methylchloroformate according to general procedure F to furnish7-chloro-1-cyclopentyl-3-ethyl-1H-pyrimido[5,4-c]quinoline-2,4-dione (15mg). LCMS: m/z 344 [M+1]⁺.

Example 311-Cyclopentyl-3,7-diethyl-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-ethyl-quinoline-3-carboxylic acid ethyl ester (250 mg, 0.95mmol) was treated with cyclopentylamine following general procedure B toafford 4-cyclopentylamino-8-ethyl-quinoline-3-carboxylic acid ethylester (250 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid in quantitative yield using general procedure D andthen transformed into the corresponding ethylamide (180 mg) followinggeneral procedure E.

The above ethylamide (0.58 mmol) was subjected to reaction with methylchloroformate according to general procedure F to furnish1-cyclopentyl-3,7-diethyl-1H-pyrimido [5,4-c]quinoline-2,4-dione (20mg). LCMS: m/z 338 [M+1]⁺.

Example 321-Cyclopentyl-3-ethyl-7-propyl-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-propyl-quinoline-3-carboxylic acid ethyl ester (250 mg, 0.9mmol) was treated with cyclopentylamine following general procedure B toafford 4-cyclopentylamino-8-propyl-quinoline-3-carboxylic acid ethylester (260 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid using general procedure D and then transformed intothe corresponding ethylamide (170 mg) following general procedure E.

The above ethylamide (0.52 mmol) was subjected to reaction with methylchloroformate according to general procedure F to furnish1-cyclopentyl-3-ethyl-7-propyl-1H-pyrimido[5,4-c]quinoline-2,4-dione (25mg). LCMS: m/z 352 [M+1]⁺.

Example 331-Cyclopentyl-3-ethyl-7-isopropyl-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-isopropyl-quinoline-3-carboxylic acid ethyl ester (250 mg,0.9 mmol) was treated with cyclopentylamine following general procedureB to afford 4-cyclopentylamino-8-isopropyl-quinoline-3-carboxylic acidethyl ester (240 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid using general procedure D and then transformed intothe corresponding ethylamide (165 mg) following general procedure E.

The above ethylamide (0.52 mmol) was subjected to reaction with methylchloroformate according to general procedure F to furnish1-cyclopentyl-3-ethyl-7-isopropyl-1H-pyrimido[5,4-c]quinoline-2,4-dione(30 mg). LCMS: m/z 352 [M+1]⁺.

Example 341-Cyclopentyl-3-ethyl-7-trifluoromethoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-trifluoromethoxy-quinoline-3-carboxylic acid ethyl ester (250mg, 0.94 mmol) was treated with cyclopentylamine following generalprocedure B to afford4-cyclopentylamino-8-trifluoromethoxy-quinoline-3-carboxylic acid ethylester (220 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid using general procedure D and then transformed intothe corresponding ethylamide (180 mg) following general procedure E. Theabove ethylamide (0.54 mmol) was subjected to reaction with methylchloroformate according to general procedure F to furnish1-cyclopentyl-3-ethyl-7-trifluoromethoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(60 mg). LCMS: m/z 394 [M+1]⁺.

Example 351-Benzyl-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg, 0.94mmol) was treated with benzylamine following general procedure B toafford 4-benzylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(220 mg). Thus obtained amino-ester was hydrolyzed to the correspondingacid using general procedure D and then transformed into thecorresponding ethylamide (180 mg) following general procedure E. Theabove ethylamide (0.54 mmol) was subjected to reaction with methylchloroformate according to general procedure F to furnish1-benzyl-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione (60mg). LCMS: m/z 362 [M+1]⁺.

Example 363-Ethyl-7-methoxy-1-(tetrahydro-pyran-4-ylmethyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (265 mg, 1.0mmol) was treated with C-(tetrahydro-pyran-4-yl)-methylamine (1.5 mmol)following general procedure B to afford8-methoxy-4-[(tetrahydro-pyran-4-ylmethyl)-amino]-quinoline-3-carboxylicacid ethyl ester (324 mg).3-Ethyl-7-methoxy-1-(tetrahydro-pyran-4-ylmethyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(19 mg) was prepared from8-methoxy-4-[(tetrahydro-pyran-4-ylmethyl)-amino]-quinoline-3-carboxylicacid ethyl ester (0.10 mmol) and ethyl isocyanate (0.4 mmol) followinggeneral procedure C. LCMS: m/z 370 [M+1]⁺.

Example 373-Ethyl-1-isopropyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (267 mg, 1.0mmol) was treated with isopropylamine (1.5 mmol) following generalprocedure B to afford 4-isopropylamino-8-methoxy-quinoline-3-carboxylicacid ethyl ester (256 mg).3-Ethyl-1-isopropyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione (14mg) was prepared from 4-isopropylamino-8-methoxy-quinoline-3-carboxylicacid ethyl ester (0.10 mmol) and ethyl isocyanate (0.4 mmol) followinggeneral procedure C. LCMS: m/z 314 [M+1]⁺.

Example 383-sec-Butyl-1-cyclopentyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-sec-Butyl-1-cyclopentyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(25 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.10 mmol) and 2-butyl isocyanate (0.4 mmol) following generalprocedure C. LCMS: m/z 368 [M+1]⁺.

Example 391-Cyclopentyl-7-methoxy-3-phenyl-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-phenyl-1H-pyrimido[5,4-c]quinoline-2,4-dione(29 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.10 mmol) and phenyl isocyanate (0.4 mmol) following general procedureC. LCMS: m/z 388 [M+1]⁺.

Example 403-Benzyl-1-cyclopentyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-Benzyl-1-cyclopentyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(28 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.10 mmol) and benzyl isocyanate (0.4 mmol) following general procedureC. LCMS: m/z 402 [M+1]⁺.

Example 411-Cyclopentyl-3-ethyl-7-methyl-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methyl-quinoline-3-carboxylic acid ethyl ester (125 mg, 0.5mmol) was treated with cyclopentylamine following general procedure B toafford 4-Cyclopentylamino-8-methyl-quinoline-3-carboxylic acid ethylester (120 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid using general procedure D and then transformed intothe corresponding ethylamide (90 mg) following general procedure E.

The above ethylamide (0.3 mmol) was subjected to reaction with methylchloroformate according to general procedure F to furnish1-cyclopentyl-3-ethyl-7-methyl-1H-pyrimido[5,4-c]quinoline-2,4-dione (10mg). LCMS: m/z 324 [M+1]⁺.

Example 42 1-Cyclopentyl-3-ethyl-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-quinoline-3-carboxylic acid ethyl ester (125 mg, 0.53 mmol) wastreated with cyclopentylamine following general procedure B to afford4-Cyclopentylamino-quinoline-3-carboxylic acid ethyl ester (110 mg).Thus obtained amino-ester was hydrolyzed to the corresponding acid usinggeneral procedure D and then transformed into the correspondingethylamide (80 mg) following general procedure E. The above ethylamide(0.28 mmol) was subjected to reaction with methyl chloroformateaccording to general procedure F to furnish1-cyclopentyl-3-ethyl-1H-pyrimido[5,4-c]quinoline-2,4-dione (15 mg).LCMS: m/z 310 [M+1]⁺.

Example 433-Ethyl-7-methoxy-1-(4-methoxy-benzyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg, 0.94mmol) was treated with 4-methoxybenzylamine following general procedureB to afford 8-methoxy-4-(4-methoxy-benzylamino)-quinoline-3-carboxylicacid ethyl ester (290 mg). Thus obtained amino-ester was hydrolyzed tothe corresponding acid using general procedure D and then transformedinto the corresponding ethylamide (190 mg) following general procedureE. The above ethylamide (0.41 mmol) was subjected to reaction withmethyl chloroformate according to general procedure F to furnish3-ethyl-7-methoxy-1-(4-methoxy-benzyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(110 mg). LCMS: m/z 422 [M+1]⁺.

Example 44 3-Ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-Ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione (25 mg) wasprepared from3-ethyl-7-methoxy-1-(4-methoxy-benzyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(50 mg, 0.11 mmol) via treatment of trifluoroacetic acid in DCM (1 mL).LCMS: m/z 272 [M+1]⁺.

Example 453-Ethyl-7-methoxy-1-(4-methyl-cyclohexyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg, 0.94mmol) was treated with 4-methyl-cyclohexylamine following generalprocedure B to afford8-methoxy-4-(4-methyl-cyclohexylamino)-quinoline-3-carboxylic acid ethylester (290 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid using general procedure D and then transformed intothe corresponding ethylamide (200 mg) following general procedure E. Theabove ethylamide (0.41 mmol) was subjected to reaction with methylchloroformate according to general procedure F to furnish3-ethyl-7-methoxy-1-(4-methyl-cyclohexyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(22 mg). LCMS: m/z 368 [M+1]⁺.

Example 46(1-Cyclopentyl-7-methoxy-2,4-dioxo-1,4-dihydro-2H-pyrimido[5,4-c]quinolin-3-yl)-aceticacid ethyl ester

(1-Cyclopentyl-7-methoxy-2,4-dioxo-1,4-dihydro-2H-pyrimido[5,4-c]quinolin-3-yl)-aceticacid ethyl ester (24 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and isocyanato-acetic acid ethyl ester (0.5 mmol) followinggeneral procedure C. LCMS: m/z 398 [M+1]⁺.

Example 473-Ethyl-1-(3-fluoro-propyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg, 0.94mmol) was treated with 3-fluoro-propylamine following general procedureB to afford 4-(3-fluoro-propylamino)-8-methoxy-quinoline-3-carboxylicacid ethyl ester (262 mg). Thus obtained amino-ester was hydrolyzed tothe corresponding acid using general procedure D and then transformedinto the corresponding ethylamide (196 mg) following general procedureE. The above ethylamide (0.64 mmol) was subjected to reaction withmethyl chloroformate according to general procedure F to furnish3-ethyl-1-(3-fluoro-propyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(138 mg). LCMS: m/z 332 [M+1]⁺.

Example 481-Cyclopentyl-7-methoxy-3-methyl-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-methyl-1H-pyrimido[5,4-c]quinoline-2,4-dione(18 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and methyl isocyanate (0.5 mmol) following general procedureC. LCMS: m/z 326 [M+1]⁺.

Example 491-Cyclopentyl-3-(3,5-dimethyl-isoxazol-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-3-(3,5-dimethyl-isoxazol-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(17 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 4-isocyanato-3,5-dimethyl-isoxazole (0.05 mmol) followinggeneral procedure C. LCMS: m/z 407 [M+1]⁺.

Example 501-Cyclopentyl-7-methoxy-3-thiophen-3-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-thiophen-3-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione(21 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 3-isocyanato-thiophene (0.5 mmol) following generalprocedure C. LCMS: m/z 394 [M+1]⁺.

Example 511-Cyclopentyl-7-methoxy-3-(4-methyl-thiophen-2-yl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-(4-methyl-thiophen-2-yl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(26 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 2-isocyanato-4-methyl-thiophene (0.5 mmol) followinggeneral procedure C. LCMS: m/z 408 [M+1]⁺. ¹H NMR (400 MHz, CDCl₃): δ9.44 (s, 1H), 8.32 (s, 1H), 7.80 (d, 1H), 7.63 (t, 1H), 7.28 (d, 1H),6.89 (s, 1H), 5.05 (p, 1H), 4.10 (s, 3H), 3.02 (s, 3H), 2.29 (m, 2H),2.11-2.19 (m, 4H), 1.25 (m, 2H) ppm.

Example 523-Ethyl-7-methoxy-1-(3-pyrrolidin-1-yl-propyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (266 mg, 1.0mmol) was treated with 3-pyrrolidin-1-yl-propylamine following generalprocedure B to afford8-methoxy-4-(3-pyrrolidin-1-yl-propylamino)-quinoline-3-carboxylic acidethyl ester (315 mg). Thus obtained amino-ester (36 mg, 0.1 mmol) wassubjected to reaction with ethyl isocyanate according to generalprocedure C to furnish3-ethyl-7-methoxy-1-(3-pyrrolidin-1-yl-propyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(19 mg). LCMS: m/z 383 [M+1]⁺.

Example 531-Cyclohexyl-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (266 mg, 1.0mmol) was treated with cyclohexylamine following general procedure B toafford 4-cyclohexylamino-8-methoxy-quinoline-3-carboxylic acid ethylester (286 mg). Thus obtained amino-ester (33 mg, 0.1 mmol) wassubjected to reaction with ethyl isocyanate according to generalprocedure C to furnish3-ethyl-7-methoxy-1-(3-pyrrolidin-1-yl-propyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(15 mg). LCMS: m/z 354 [M+1]⁺. ¹H NMR (400 MHz, CDCl₃): δ 9.58 (s, 1H),7.58 (m, 1H), 7.50 (m, 1H), 7.41 (m, 1H), 7.24 (m, 1H), 7.06-7.14 (m,2H), 6.48 (m, 1H), 4.92 (m, 1H), 4.08 (s, 3H), 2.46 (m, 2H), 1.64-1.92(m, 5H), 1.42 (m, 2H), 1.28 (m, 1H) ppm.

Example 541-Cyclopentyl-7-methoxy-3-m-tolyl-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-m-tolyl-1H-pyrimido[5,4-c]quinoline-2,4-dione(29 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 1-isocyanato-3-methyl-benzene (0.5 mmol) followinggeneral procedure C. LCMS: m/z 402 [M+1]⁺. ¹H NMR (400 MHz, CDCl₃): δ9.46 (s, 1H), 7.77 (d, 1H), 7.53 (t, 1H), 7.46 (d, 1H), 7.32 (s, 1H),7.16 (d, 1H), 7.14 (d, 1H), 6.83 (d, 1H), 5.06 (p, 1H), 4.12 (s, 3H),2.41 (m, 2H), 2.32 (s, 3H), 1.98 (m, 4H), 1.32 (m, 2H) ppm.

Example 551-Cyclopentyl-7-methoxy-3-p-tolyl-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-p-tolyl-1H-pyrimido[5,4-c]quinoline-2,4-dione(26 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 1-isocyanato-4-methyl-benzene (0.5 mmol) followinggeneral procedure C. LCMS: m/z 402 [M+1]⁺. ¹H NMR (400 MHz, CDCl₃): δ9.49 (s, 1H), 7.84 (d, 1H), 7.61 (t, 1H), 7.38 (m, 1H), 7.34 (m, 1H),7.32 (m, 1H), 7.24 (d, 1H), 7.08 (d, 1H), 5.03 (p, 1H), 4.16 (s, 3H),2.43 (m, 2H), 2.29 (s, 3H), 2.08 (m, 4H), and 1.24 (m, 2H) ppm.

Example 568-Chloro-3-ethyl-1-(4-fluoro-butyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4,7-Dichloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg,84 mmol) was treated with 4-fluoro-butylamine following generalprocedure B to afford7-chloro-4-(4-fluoro-butylamino)-8-methoxy-quinoline-3-carboxylic acidethyl ester (240 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid using general procedure D and then transformed intothe corresponding ethylamide (175 mg) following general procedure E. Theabove ethylamide (0.52 mmol) was subjected to reaction with methylchloroformate according to general procedure F to furnish8-chloro-3-ethyl-1-(4-fluoro-butyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(117 mg). LCMS: m/z 380 [M+1]⁺.

Example 571-Butyl-3-ethyl-9-fluoro-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-6-fluoro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250mg, 0.88 mmol) was treated with butylamine following general procedure Bto afford 4-butylamino-6-fluoro-8-methoxy-quinoline-3-carboxylic acidethyl ester (282 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid using general procedure D and then transformed intothe corresponding ethylamide (170 mg) following general procedure E. Theabove ethylamide (0.170 mmol) was subjected to reaction with methylchloroformate according to general procedure F to furnish1-butyl-3-ethyl-9-fluoro-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(105 mg). LCMS: m/z 346 [M+1]⁺.

Example 581-Butyl-9-chloro-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4,6-Dichloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg,0.84 mmol) was treated with butylamine following general procedure B toafford 4-butylamino-6-chloro-8-methoxy-quinoline-3-carboxylic acid ethylester (230 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid using general procedure D and then transformed intothe corresponding ethylamide (155 mg) following general procedure E. Theabove ethylamide (155 mg, 0.45 mmol) was subjected to reaction withmethyl chloroformate according to general procedure F to furnish1-butyl-9-chloro-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(87 mg). LCMS: m/z 362 [M+1]⁺.

Example 599-Chloro-3-ethyl-1-(4-fluoro-butyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4,6-Dichloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg,0.836 mmol) was treated with 4-fluorobutylamine following generalprocedure B to afford6-Chloro-4-(4-fluoro-butylamino)-8-methoxy-quinoline-3-carboxylic acidethyl ester (185 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid using general procedure D and then transformed intothe corresponding ethylamide (105 mg) following general procedure E. Theabove ethylamide (100 mg, 0.28 mmol) was subjected to reaction withmethyl chloroformate according to general procedure F to furnish9-chloro-3-ethyl-1-(4-fluoro-butyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(54 mg). LCMS: m/z 380 [M+1]⁺.

Example 601-Butyl-8-chloro-3-ethyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4,7-Dichloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg,0.84 mmol) was treated with butylamine following general procedure B toafford 7-chloro-4-butylamino-8-methoxy-quinoline-3-carboxylic acid ethylester (210 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid using general procedure D and then transformed intothe corresponding ethylamide (125 mg) following general procedure E. Theabove ethylamide (0.37 mmol) was subjected to reaction with methylchloroformate according to general procedure F to furnish8-chloro-3-ethyl-1-butyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(75 mg). LCMS: m/z 380 [M+1]⁺.

Example 613-Ethyl-9-fluoro-1-(4-fluoro-butyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-6-fluoro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250mg, 0.88 mmol) was treated with 4-fluoro-butylamine following generalprocedure B to afford6-fluoro-4-(4-fluoro-butylamino)-8-methoxy-quinoline-3-carboxylic acidethyl ester (175 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid using general procedure D and then transformed intothe corresponding ethylamide (95 mg) following general procedure E. Theabove ethylamide (95 mg, 0.28 mmol) was subjected to reaction withmethyl chloroformate according to general procedure F to furnish3-ethyl-9-fluoro-1-(4-fluoro-butyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(52 mg). LCMS: m/z 364 [M+1]⁺.

Example 623-Ethyl-1-(4-fluoro-butyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg, 0.94mmol) was treated with 4-fluoro-butylamine following general procedure Bto afford 4-(4-fluoro-butylamino)-8-methoxy-quinoline-3-carboxylic acidethyl ester (244 mg). Thus obtained amino-ester was hydrolyzed to thecorresponding acid using general procedure D and then transformed intothe corresponding ethylamide (150 mg) following general procedure E. Theabove ethylamide (145 mg, 0.45 mmol) was subjected to reaction withmethyl chloroformate according to general procedure F to furnish3-ethyl-1-(4-fluoro-butyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(95 mg). LCMS: m/z 346 [M+1]⁺.

Example 631-Cyclopentyl-3-(4-fluoro-3-methyl-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-3-(4-fluoro-3-methyl-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(25 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 1-fluoro-4-isocyanato-2-methyl-benzene (0.5 mmol)following general procedure C. LCMS: m/z 420 [M+1]⁺.

Example 641-Cyclopentyl-3-(3-fluoro-4-methyl-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-3-(3-fluoro-4-methyl-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(28 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 1-fluoro-5-isocyanato-2-methyl-benzene (0.5 mmol)following general procedure C. LCMS: m/z 420 [M+1]⁺.

Example 651-Cyclopentyl-7-methoxy-3-(3-methoxy-phenyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-(3-methoxy-phenyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(32 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 1-isocyanato-3-methoxy-benzene (0.5 mmol) followinggeneral procedure C. LCMS: m/z 418 [M+1]⁺. ¹H NMR (400 MHz, CDCl₃): δ9.50 (s, 1H), 7.86 (d, 1H), 7.64 (t, 1H), 7.44 (t, 1H), 7.30 (d, 1H),6.82 (dd, 1H), 6.60 (d, 1H), 6.28 (s, 1H), 5.04 (p, 1H), 4.17 (s, 3H),3.80 (s, 3H), 2.40 (m, 2H), 2.04-2.20 (m, 4H), 1.24 (m, 2H) ppm.

Example 661-Cyclopentyl-3-(3-fluoro-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-3-(3-fluoro-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(23 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 1-fluoro-3-isocyanato-benzene (0.5 mmol) followinggeneral procedure C. LCMS: m/z 406 [M+1]⁺. ¹H NMR (400 MHz, CDCl₃): δ9.45 (s, 1H), 7.84 (d, 1H), 7.62 (d, 1H), 7.51 (t, 1H), 7.20 (m, 1H),7.18 (m, 1H), 7.12 (m, 1H), 7.09 (s, 1H), 5.06 (p, 1H), 4.15 (s, 3H),2.42 (m, 2H), 2.06-2.18 (m, 4H), 1.26 (m, 2H) ppm.

Example 671-Cyclopentyl-3-(4-difluoromethoxy-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-3-(4-difluoromethoxy-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(19 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 1-difluoromethoxy-4-isocyanato-benzene (0.5 mmol)following general procedure C. LCMS: m/z 454 [M+1]⁺.

Example 681-Cyclopentyl-7-methoxy-3-(4-trifluoromethoxy-phenyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-(4-trifluoromethoxy-phenyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(24 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 1-isocyanato-4-trifluoromethoxy-benzene (0.5 mmol)following general procedure C. LCMS: m/z 472 [M+1]⁺.

Example 691-Cyclopentyl-7-methoxy-3-(2-trifluoromethyl-phenyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-(2-trifluoromethyl-phenyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(21 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 1-isocyanato-2-trifluoromethyl-benzene (0.5 mmol)following general procedure C. LCMS: m/z 456 [M+1]⁺.

Example 701-Cyclopentyl-7-methoxy-3-(5-methyl-2-trifluoromethyl-furan-3-yl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-(5-methyl-2-trifluoromethyl-furan-3-yl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(25 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 3-isocyanato-5-methyl-2-trifluoromethyl-furan (0.5 mmol)following general procedure C. LCMS: m/z 460 [M+1]⁺.

Example 711-Cyclopentyl-7-methoxy-3-(3-methyl-benzyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-(3-methyl-benzyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(18 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 1-isocyanatomethyl-3-methyl-benzene (0.5 mmol) followinggeneral procedure C. LCMS: m/z 416 [M+1]⁺.

Example 723-(3-Chloro-phenyl)-7-methoxy-1-(4-methyl-cyclohexyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-(3-Chloro-phenyl)-7-methoxy-1-(4-methyl-cyclohexyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(115 mg) was prepared from8-methoxy-4-(4-methyl-cyclohexylamino)-quinoline-3-carboxylic acid ethylester (150 mg, 0.44 mmol) and 1-chloro-3-isocyanato-benzene (0.66 mmol)following general procedure C. LCMS: m/z 450 [M+1]⁺.

Example 733-(3-Chloro-phenyl)-1-cyclopentyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-(3-Chloro-phenyl)-1-cyclopentyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(55 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.48 mmol) and 1-chloro-3-isocyanato-benzene (0.72 mmol) followinggeneral procedure C. LCMS: m/z 422 [M+1]⁺.

Example 741-Cyclopentyl-3-(3,5-dimethyl-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-3-(3,5-dimethyl-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(33 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.32 mmol) and 1-isocyanato-3,5-dimethyl-benzene (0.48 mmol) followinggeneral procedure C. LCMS: m/z 416 [M+1]⁺.

Example 751-Cyclopentyl-3-(3-ethyl-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-3-(3-ethyl-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(89 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.32 mmol) and 1-ethyl-3-isocyanato-benzene (0.48 mmol) followinggeneral procedure C. LCMS: m/z 416 [M+1]⁺.

Example 761-Cyclopentyl-3-(2,3-dimethyl-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-3-(2,3-dimethyl-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(22 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.32 mmol) and 1-isocyanato-2,3-dimethyl-benzene (0.48 mmol) followinggeneral procedure C. LCMS: m/z 416 [M+1]⁺.

Example 771-Cyclopentyl-3-(2,5-dimethyl-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-3-(2,5-dimethyl-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(46 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.32 mmol) and 1-isocyanato-2,5-dimethyl-benzene (0.48 mmol) followinggeneral procedure C. LCMS: m/z 416 [M+1]⁺.

Example 787-Methoxy-1-(4-methyl-cyclohexyl)-3-m-tolyl-1H-pyrimido[5,4-c]quinoline-2,4-dione

7-Methoxy-1-(4-methyl-cyclohexyl)-3-m-tolyl-1H-pyrimido[5,4-c]quinoline-2,4-dione(135 mg) was prepared from8-methoxy-4-(4-methyl-cyclohexylamino)-quinoline-3-carboxylic acid ethylester (0.29 mmol) and 1-isocyanato-3-methyl-benzene (0.44 mmol)following general procedure C. LCMS: m/z 430 [M+1]⁺.

Example 791-Cyclopentyl-7-methoxy-3-(3-nitro-phenyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-(3-nitro-phenyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(19 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 1-isocyanato-3-nitro-benzene (0.5 mmol) following generalprocedure C. LCMS: m/z 433 [M+1]⁺.

Example 801-Cyclopentyl-7-methoxy-3-(3-trifluoromethyl-phenyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-(3-trifluoromethyl-phenyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(27 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 1-isocyanato-3-nitro-benzene (0.5 mmol) following generalprocedure C. LCMS: m/z 456 [M+1]⁺.

Example 811-Cyclopentyl-3-(3,4-dimethyl-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-3-(3,4-dimethyl-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(106 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.32 mmol) and 1-isocyanato-3,4-dimethyl-benzene (0.48 mmol) followinggeneral procedure C. LCMS: m/z 416 [M+1]⁺.

Example 821-Cyclopentyl-7-methoxy-3-o-tolyl-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-o-tolyl-1H-pyrimido[5,4-c]quinoline-2,4-dione(24 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 1-isocyanato-2-methyl-benzene (0.5 mmol) followinggeneral procedure C. LCMS: m/z 402 [M+1]⁺.

Example 831-Butyl-7-methoxy-3-m-tolyl-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Butyl-7-methoxy-3-m-tolyl-1H-pyrimido[5,4-c]quinoline-2,4-dione (17mg) was prepared from 4-butylamino-8-methoxy-quinoline-3-carboxylic acidethyl ester (0.1 mmol) and 1-isocyanato-3-methyl-benzene (0.5 mmol)following general procedure C. LCMS: m/z 390 [M+1]⁺.

Example 843-Benzo[1,3]dioxol-5-yl-1-cyclopentyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-Benzo[1,3]dioxol-5-yl-1-cyclopentyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(33 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 5-isocyanato-benzo[1,3]dioxole (0.5 mmol) followinggeneral procedure C. LCMS: m/z 432 [M+1]⁺.

Example 851-Cyclopentyl-3-indan-5-yl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-3-indan-5-yl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(21 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 5-isocyanato-indan (0.5 mmol) following general procedureC. LCMS: m/z 428 [M+1]⁺.

Example 861-Cyclopentyl-7-methoxy-3-phenethyl-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-phenethyl-1H-pyrimido[5,4-c]quinoline-2,4-dione(20 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and (2-phenyl)ethyl isocyanate (0.5 mmol) following generalprocedure C. LCMS: m/z 416 [M+1]⁺.

Example 873-(3-Chloro-phenyl)-1-(4,4-difluoro-cyclohexyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg, 0.94mmol) was treated with 4,4-difluoro-cyclohexylamine following generalprocedure B to afford4-(4,4-difluoro-cyclohexylamino)-8-methoxy-quinoline-3-carboxylic acidethyl ester (192 mg). Thus obtained amino-ester (92 mg, 0.25 mmol) wassubjected to reaction with 1-chloro-3-isocyanato-benzene according togeneral procedure C to furnish3-(3-chloro-phenyl)-1-(4,4-difluoro-cyclohexyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(69 mg). LCMS: m/z 472 [M+1]⁺. ¹H NMR (400 MHz, CDCl₃): δ 9.56 (s, 1H),7.58 (m, 1H), 7.48 (m, 1H), 7.38 (m, 1H), 7.24 (m, 1H), 7.06-7.14 (m,2H), 6.48 (m, 1H), 5.09 (m, 1H), 4.08 (s, 3H), 2.84 (m, 2H), 2.23 (m,2H), 1.80-2.00 (m, 4H) ppm.

Example 884-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-piperidine-1-carboxylicacid tert-butyl ester

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (1.0 g, 3.77mmol) was treated with 4-amino-piperidine-1-carboxylic acid tert-butylester following general procedure B to afford4-(1-tert-butoxycarbonyl-piperidin-4-ylamino)-8-methoxy-quinoline-3-carboxylicacid ethyl ester (1.48 g). Thus obtained amino-ester (500 mg, 1.17 mmol)was subjected to reaction with 1-chloro-3-isocyanato-benzene accordingto general procedure C to furnish4-[3-(3-chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-piperidine-1-carboxylicacid tert-butyl ester (500 mg). LCMS: m/z 537 [M+1]⁺.

Example 893-(3-Chloro-phenyl)-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochlode

3-(3-Chloro-phenyl)-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(415 mg) was prepared according to general procedure G from4-[3-(3-chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-piperidine-1-carboxylicacid tert-butyl ester (475 mg, 0.89 mmol). LCMS: m/z 437 [M+1]⁺.

Example 901-(1-Acetyl-piperidin-4-yl)-3-(3-chloro-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-(1-Acetyl-piperidin-4-yl)-3-(3-chloro-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(45 mg) was prepared according to general procedure H from3-(3-chloro-phenyl)-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(50 mg, 0.1 mmol) and acetyl chloride. LCMS: m/z 479 [M+1]⁺. ¹H NMR (400MHz, CDCl₃): δ 9.59 (s, 1H), 7.58 (m, 1H), 7.52 (m, 1H), 7.42 (m, 1H),7.24 (m, 1H), 7.10-7.18 (m, 2H), 6.48 (m, 1H), 5.18 (m, 1H), 4.82 (m,1H), 4.12 (s, 3H), 3.95 (m, 1H), 3.20 (m, 1H), 3.58-3.78 (m, 3H), 2.08(s, 3H), 1.78 (m, 2H) ppm.

Example 911-(1-Benzoyl-piperidin-4-yl)-3-(3-chloro-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-(1-Benzoyl-piperidin-4-yl)-3-(3-chloro-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(51 mg) was prepared according to general procedure H from3-(3-chloro-phenyl)-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(50 mg, 0.1 mmol) and benzoyl chloride. LCMS: m/z 541 [M+1]⁺.

Example 923-(3-Chloro-phenyl)-1-(1-methanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-(3-Chloro-phenyl)-1-(1-methanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(47 mg) was prepared according to general procedure H from3-(3-chloro-phenyl)-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(50 mg, 0.1 mmol) and methanesulfonyl chloride. LCMS: m/z 515 [M+1]⁺. ¹HNMR (400 MHz, CDCl₃): δ 9.56 (s, 1H), 7.58 (m, 1H), 7.48 (m, 1H), 7.38(m, 1H), 7.24 (m, 1H), 7.09-7.18 (m, 2H), 6.48 (m, 1H), 5.06 (m, 1H),4.10 (s, 3H), 4.02 (m, 2H), 3.80-3.92 (m, 7H), 1.84 (m, 2H) ppm.

Example 933-(3-Chloro-phenyl)-7-methoxy-1-[1-(propane-2-sulfonyl)-piperidin-4-yl]-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-(3-Chloro-phenyl)-7-methoxy-1-[1-(propane-2-sulfonyl)-piperidin-4-yl]-1H-pyrimido[5,4-c]quinoline-2,4-dione(51 mg) was prepared according to general procedure H from3-(3-chloro-phenyl)-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(50 mg, 0.1 mmol) and methanesulfonyl chloride. LCMS: m/z 543 [M+1]⁺.

Example 944-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-piperidine-1-sulfonicacid dimethylamide

4-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-piperidine-1-sulfonicacid dimethylamide (48 mg) was prepared according to general procedure Hfrom3-(3-chloro-phenyl)-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(50 mg, 0.1 mmol) and N,N-dimethylsulfamoyl chloride. LCMS: m/z 544[M+1]⁺.

Example 951-(1-Benzenesulfonyl-piperidin-4-yl)-3-(3-chloro-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-(1-Benzenesulfonyl-piperidin-4-yl)-3-(3-chloro-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(51 mg) was prepared according to general procedure H from3-(3-chloro-phenyl)-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(50 mg, 0.1 mmol) and benzenesulfonyl chloride. LCMS: m/z 577 [M+1]⁺.

Example 963-(3-Chloro-phenyl)-1-[1-(1,1-dioxo-tetrahydro-1lambda*6*-thiophene-3-sulfonyl)-piperidin-4-yl]-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-(3-Chloro-phenyl)-1-[1-(1,1-dioxo-tetrahydro-thiophene-3-sulfonyl)-piperidin-4-yl]-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(51 mg) was prepared according to general procedure H from3-(3-chloro-phenyl)-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(50 mg, 0.1 mmol) and 1,1-dioxo-tetrahydro-thiophene-3-sulfonylchloride. LCMS: m/z 619 [M+1]⁺.

Example 974-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-piperidine-1-carboxylicacid ethylamide

4-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-piperidine-1-carboxylicacid ethylamide (40 mg) was prepared according to general procedure Hfrom3-(3-chloro-phenyl)-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(50 mg, 0.1 mmol) and ethyl isocyanate. LCMS: m/z 508 [M+1]⁺.

Example 984-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-piperidine-1-carboxylicacid isopropylamide

4-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-piperidine-1-carboxylicacid isopropylamide (39 mg) was prepared according to general procedureH from3-(3-chloro-phenyl)-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(50 mg, 0.10 mmol) and isopropyl isocyanate. LCMS: m/z 508 [M+1]⁺.

Example 993-(2-Chloro-phenyl)-7-methoxy-1-(4-methyl-cyclohexyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-(2-Chloro-phenyl)-7-methoxy-1-(4-methyl-cyclohexyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(50 mg) was prepared from8-methoxy-4-(4-methyl-cyclohexylamino)-quinoline-3-carboxylic acid ethylester (0.164 mmol) and 1-chloro-2-isocyanato-benzene (0.25 mmol)following general procedure C. LCMS: m/z 450 [M+1]⁺. ¹H NMR (400 MHz,CDCl₃): δ 9.58 (s, 1H), 7.62 (m, 1H), 7.54-7.59 (m, 1H), 7.52 (m, 2H),7.06 (m, 2H), 6.42 (m, 1H), 4.91 (m, 1H), 4.06 (s, 3H), 2.71 (m, 1H),1.98 (m, 1H), 1.65 (m, 4H), 1.56 (m, 1H), 1.08 (d, 3H) ppm.

Example 1003-(4-Chloro-phenyl)-7-methoxy-1-(4-methyl-cyclohexyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-(4-Chloro-phenyl)-7-methoxy-1-(4-methyl-cyclohexyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(55 mg) was prepared from8-methoxy-4-(4-methyl-cyclohexylamino)-quinoline-3-carboxylic acid ethylester (0.164 mmol) and 1-chloro-4-isocyanato-benzene (0.25 mmol)following general procedure C. LCMS: m/z 450 [M+1]⁺.

Example 1014-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-piperidine-1-carboxylicacid isopropyl ester

4-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-piperidine-1-carboxylicacid isopropyl ester (47 mg) was prepared according to general procedureH from3-(3-chloro-phenyl)-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(50 mg, 0.105 mmol) and isopropyl chloroformate. LCMS: m/z 523 [M+1]⁺.

Example 102Trans-4-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-cyclohexane-carboxylicacid methyl ester

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg, 0.94mmol) was treated with trans-4-amino-cyclohexanecarboxylic acidethylester following general procedure B to afford8-methoxy-4-(trans-4-ethoxycarbonyl-cyclohexylamino)-quinoline-3-carboxylicacid ethyl ester (207 mg). Thus obtained amino-ester (200 mg, 0.50 mmol)was subjected to reaction with 1-chloro-3-isocyanato-benzene accordingto general procedure C to furnishtrans-4-[3-(3-chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-cyclohexane-carboxylicacid ethyl ester (175 mg). LCMS: m/z 508 [M+1]⁺.

Example 103Trans-4-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-cyclohexanecarboxylicacid.hydrochloride

Trans-4-[3-(3-chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-cyclohexanecarboxylicacid.hydrochloride (31 mg) was prepared according to general procedure Ifromtrans-4-[3-(3-chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-cyclohexane-carboxylicacid ethyl ester (75 mg, 0.15 mmol). LCMS: m/z 480 [M+1]⁺.

Example 104Cis-4-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-cyclohexane-carboxylicacid ethyl ester

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (250 mg, 0.94mmol) was treated with cis-4-amino-cyclohexanecarboxylic acid ethylester following general procedure B to afford8-methoxy-4-(cis-4-ethoxycarbonyl-cyclohexylamino)-quinoline-3-carboxylicacid ethyl ester (198 mg). Thus obtained amino-ester (175 mg, 0.44 mmol)was subjected to reaction with 1-chloro-3-isocyanato-benzene accordingto general procedure C to furnishcis-4-[3-(3-chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-cyclohexane-carboxylicacid ethyl ester (138 mg). LCMS: m/z 508 [M+1]⁺.

Example 105Cis-4-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-cyclohexane-carboxylicacid.hydrochloride

Cis-4-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-cyclohexanecarboxylicacid.hydrochloride (35 mg) was prepared according to general procedure Ifromcis-4-[3-(3-chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-cyclohexane-carboxylicacid ethyl ester (75 mg, 0.15 mmol). LCMS: m/z 480 [M+1]⁺.

Example 1063-(3-Chloro-phenyl)-1-cyclohexyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-(3-Chloro-phenyl)-1-cyclohexyl-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(30 mg) was prepared from4-cyclohexylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester (0.1mmol) and 1-chloro-3-isocyanato-benzene (0.15 mmol) following generalprocedure C. LCMS: m/z 436 [M+1]⁺.

Example 1074-(3-Benzo[1,3]dioxol-5-yl-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl)-piperidine-1-carboxylicacid tert-butyl ester

4-(3-Benzo[1,3]dioxol-5-yl-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl)-piperidine-1-carboxylicacid tert-butyl ester (110 mg) was prepared from4-(1-tert-butoxycarbonyl-piperidin-4-ylamino)-8-methoxy-quinoline-3-carboxylicacid ethyl ester (0.31 mmol) and 5-isocyanato-benzo[1,3]dioxole (0.465mmol) following general procedure C. LCMS: m/z 547 [M+1]⁺.

Example 1083-Benzo[1,3]dioxol-5-yl-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride

3-Benzo[1,3]dioxol-5-yl-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(88 mg) was prepared according to general procedure G from4-(3-benzo[1,3]dioxol-5-yl-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl)-piperidine-1-carboxylicacid tert-butyl ester (100 mg, 183 mmol). LCMS: m/z 447 [M+1]⁺.

Example 1093-Benzo[1,3]dioxol-5-yl-1-(1-methanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-Benzo[1,3]dioxol-5-yl-1-(1-methanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(45 mg) was prepared according to general procedure H from3-Benzo[1,3]dioxol-5-yl-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(50 mg, 0.1 mmol) and methanesulfonyl chloride. LCMS: m/z 525 [M+1]⁺. ¹HNMR (400 MHz, CDCl₃): δ 9.52 (s, 1H), 7.15 (m, 1H), 7.08 (m, 1H), 6.92(m, 1H), 6.86 (m, 1H), 6.70-6.76 (m, 2H), 6.19 (s, 1H), 6.12 (s, 1H),5.09 (m, 1H), 4.38 (m, 1H), 4.18 (m, 2H), 4.07 (s, 3H), 3.26 (m, 2H),2.91 (m, 2H), 2.68 (m, 2H), 1.78 (m, 2H), 1.14 (t, 3H) ppm.

Example 1103-(1-Cyclopentyl-7-methoxy-2,4-dioxo-1,4-dihydro-2H-pyrimido[5,4-c]quinolin-3-yl)-benzonitrile

3-(1-Cyclopentyl-7-methoxy-2,4-dioxo-1,4-dihydro-2H-pyrimido[5,4-c]quinolin-3-yl)-benzonitrile(19 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 3-isocyanato-benzonitrile (0.5 mmol) following generalprocedure C. LCMS: m/z 413 [M+1]⁺.

Example 1111-Cyclopentyl-7-methoxy-3-(3-methylsulfanyl-phenyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-7-methoxy-3-(3-methylsulfanyl-phenyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(28 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 1-isocyanato-3-methylsulfanyl-benzene (0.5 mmol)following general procedure C. LCMS: m/z 434 [M+1]⁺.

Example 1121-Cyclopentyl-3-(3-methanesulfonyl-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-3-(3-methanesulfonyl-phenyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(21 mg) was prepared according to general procedure J from1-cyclopentyl-7-methoxy-3-(3-methylsulfanyl-phenyl)-1H-pyrimido[5,4-c]quinoline-2,4-dione(21 mg, 0.5 mmol). LCMS: m/z 466 [M+1]⁺.

Example 1133-[(R)-3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-pyrrolidine-1-carboxylicacid tert-butyl ester

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (266 mg, 1.0mmol) was treated with (R)-3-amino-pyrrolidine-1-carboxylic acidtert-butyl ester following general procedure B to afford4-((R)-1-tert-butoxycarbonyl-pyrrolidin-3-ylamino)-8-methoxy-quinoline-3-carboxylicacid ethyl ester 381 mg). Thus obtained amino-ester (208 mg, 0.5 mmol)was subjected to reaction with 1-chloro-3-isocyanato-benzene accordingto general procedure C to furnish3-[(R)-3-(3-chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-pyrrolidine-1-carboxylicacid tert-butyl ester (194 mg). LCMS: m/z 523 [M+1]⁺.

Example 114(R)-3-(3-Chloro-phenyl)-7-methoxy-1-pyrrolidin-3-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride

(R)-3-(3-Chloro-phenyl)-7-methoxy-1-pyrrolidin-3-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(59 mg) was prepared according to general procedure G from3-[(R)-3-(3-chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-pyrrolidine-1-carboxylicacid tert-butyl ester (105 mg, 0.2 mmol). LCMS: m/z 423 [M+1]⁺.

Example 1153-(3-Chloro-phenyl)-1-((R)-1-methanesulfonyl-pyrrolidin-3-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-(3-Chloro-phenyl)-1-((R)-1-methanesulfonyl-pyrrolidin-3-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(29 mg) was prepared according to general procedure H from(R)-3-(3-chloro-phenyl)-7-methoxy-1-pyrrolidin-3-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(42 mg, 0.1 mmol) and methanesulfonyl chloride. LCMS: m/z 501 [M+1]⁺.

Example 1163-[(S)-3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-pyrrolidine-1-carboxylicacid tert-butyl ester

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (266 mg, 1.0mmol) was treated with (S)-3-amino-pyrrolidine-1-carboxylic acidtert-butyl ester following general procedure B to afford4-((S)-1-tert-butoxycarbonyl-pyrrolidin-3-ylamino)-8-methoxy-quinoline-3-carboxylicacid ethyl ester (345 mg). Thus obtained amino-ester (208 mg, 0.5 mmol)was subjected to reaction with 1-chloro-3-isocyanato-benzene accordingto general procedure C to furnish3-[(S)-3-(3-chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-pyrrolidine-1-carboxylicacid tert-butyl ester (177 mg). LCMS: m/z 523 [M+1]⁺.

Example 117(S)-3-(3-Chloro-phenyl)-7-methoxy-1-pyrrolidin-3-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride

(S)-3-(3-Chloro-phenyl)-7-methoxy-1-pyrrolidin-3-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(53 mg) was prepared according to general procedure G from3-[(S)-3-(3-chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-pyrrolidine-1-carboxylicacid tert-butyl ester (105 mg, 0.2 mmol). LCMS: m/z 423 [M+1]⁺.

Example 1183-(3-Chloro-phenyl)-1-((S)-1-methanesulfonyl-pyrrolidin-3-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-(3-Chloro-phenyl)-1-((S)-1-methanesulfonyl-pyrrolidin-3-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(27 mg) was prepared according to general procedure H from(S)-3-(3-chloro-phenyl)-7-methoxy-1-pyrrolidin-3-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(42 mg, 0.1 mmol) and methanesulfonyl chloride. LCMS: m/z 501 [M+1]⁺.

Example 1193-Benzo[1,3]dioxol-5-yl-1-(2-methanesulfonyl-ethyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

4-Chloro-8-methoxy-quinoline-3-carboxylic acid ethyl ester (266 mg, 1.0mmol) was treated with 2-methanesulfonyl-ethylamine following generalprocedure B to afford4-(2-methanesulfonyl-ethylamino)-8-methoxy-quinoline-3-carboxylic acidethyl ester (305 mg). Thus obtained amino-ester (70 mg, 0.2 mmol) wassubjected to reaction with 5-Isocyanato-benzo[1,3]dioxole according togeneral procedure C to furnish3-Benzo[1,3]dioxol-5-yl-1-(2-methanesulfonyl-ethyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(43 mg). LCMS: m/z 471 [M+1]⁺.

Example 1201-Cyclopentyl-3-(2,3-dihydro-benzo[1,4]dioxin-6-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

1-Cyclopentyl-3-(2,3-dihydro-benzo[1,4]dioxin-6-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(18 mg) was prepared from4-cyclopentylamino-8-methoxy-quinoline-3-carboxylic acid ethyl ester(0.1 mmol) and 6-isocyanato-2,3-dihydro-benzo[1,4]dioxine (0.5 mmol)following general procedure C. LCMS: m/z 446 [M+1]⁺.

Example 1213-(3-Chloro-phenyl)-1-(1-ethanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-(3-Chloro-phenyl)-1-(1-ethanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(32 mg) was prepared according to general procedure H from3-(3-chloro-phenyl)-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(40 mg) and ethanesulfonyl chloride. LCMS: m/z 529 [M+1]⁺.

Example 1223-(3-Chloro-phenyl)-1-(1-methanesulfonylmethanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

3-(3-Chloro-phenyl)-1-(1-ethanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(42 mg) was prepared according to general procedure H from3-(3-chloro-phenyl)-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(40 mg) and methanesulfonylmethanesulfonyl chloride. LCMS: m/z 594[M+1]⁺.

Example 1234-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-piperidine-1-carboxylicacid methylamide

3-(3-Chloro-phenyl)-1-(1-ethanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione(30 mg) was prepared by reacting3-(3-chloro-phenyl)-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(75 mg) with carbonyldiimidazole and methylamine. LCMS: m/z 494 [M+1]⁺.

Example 1244-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-piperidine-1-carboxylicacid methyl ester

4-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-piperidine-1-carboxylicacid methyl ester (35 mg) was prepared by reacting3-(3-chloro-phenyl)-7-methoxy-1-piperidin-4-yl-1H-pyrimido[5,4-c]quinoline-2,4-dione.dihydrochloride(40 mg) with carbonyldiimidazole and methylamine. LCMS: m/z 495 [M+1]⁺.

Example 1253-(3-2Chloro-phenyl)-1-(1-methanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

Title compound was prepared using the same procedures described for thepreparation of3-(3-chloro-phenyl)-1-(1-methanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione.LCMS: m/z 515 [M+1]⁺.

Example 1263-(3-Chloro-phenyl)-1-(1-methanesulfonyl-piperidin-4-ylmethyl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione

Title compound was prepared using the same procedures described for thepreparation of3-(3-chloro-phenyl)-1-(1-methanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dioneusing 4-aminomethyl-piperidine-1-carboxylic acid tert-butyl ester. LCMS:m/z 529 [M+1]⁺.

Example 127N-{4-[(S)-3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-trans-cyclohexyl}-methanesulfonamide

Title compound was prepared using the same procedures described for thepreparation of3-(3-chloro-phenyl)-1-(1-methanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dioneusing trans-(4-amino-cyclohexyl)-carbamic acid tert-butyl ester exceptthe cyclization was carried out following general procedure K. LCMS: m/z529 [M+1]⁺.

Example 128N-{3-[3-(3-Chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-propyl}-methanesulfonamide

Title compound was prepared using the same procedures described for thepreparation of3-(3-chloro-phenyl)-1-(1-methanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dioneexcept that the cyclization of amine-ester was carried out followinggeneral procedure K. LCMS: m/z 489 [M+1]⁺.

Biological Examples

The compounds of the present invention include inhibitors of TNF-αsynthesis. Compounds can be assayed in vitro for their ability toinhibit TNF-α accumulation in cell cultures. Inhibitor binding may alsobe measured by radiolabelling the inhibitor prior to binding, isolatingthe inhibitor or complex and determining the amount of radiolabel bound.Alternatively, inhibitor binding may be determined by running acompetition experiment in which new inhibitors are incubated with aknown radioligand. Compounds also can be assayed for their ability toaffect cellular or physiological functions mediated by activity. Assaysfor each of these activities are described herein or are known in theart.

In general, embodiments of the present invention useful forpharmaceutical applications may have inhibitory potencies (IC₅₀'s) ofinterest of below about 100 μM. In an embodiment, embodiments of thepresent invention useful for pharmaceutical applications may have anIC₅₀ for a protein of interest of below about 50 μM. For particularapplications, lower inhibitory potencies may be useful. Thus, in anotherembodiment, compounds of the present invention may inhibit TNF-α with anIC₅₀ in a range of less than 10 μM.

The compounds of the present invention elicit important and measurablepharmacological responses.

Human Whole Blood Assay

Blood is obtained by venipuncture from a healthy human donor. Preferablyall donors are pre-screened for sexually transmitted disease as well asHIV and have been given a physical by a physician. Blood is drawn into50 cc syringes containing 0.5 ml of sodium heparin (BD, 5000 U/mL).Typically each donor will give 100-150 ml of heparinized blood.

A third 50 cc syringe is collected without heparin. The non-heparinzedblood is immediately evacuated into 5 to 6-8 ml clotting tubes (red-graycap) containing a gel plug. (Becton Dickinson).

The clotted blood is separated by centrifugation at 1200×g for 15minutes. The serum is removed, placed into a sterile tube and reserved.

Compounds are prepared as DMSO stocks. All compounds are prepared as 20mM solutions and then serial diluted with DMSO. The final DMSOconcentrations are not greater than 0.5% in the assay.

Compounds are added to a sterile deep-well polypropylene block (Corning)followed by 360 μL of heparinized blood. The blocks are covered with anadhesive foil strip then inverted to mix.

The blocks containing the compound and blood are placed into a 37° C.incubator for 30 minutes.

A stock of LPS using the reserved human serum is prepared. To preparethe stock, add 1 μL of LPS (5 mg/mL; Sigma Cat #L3024) to 5 mL of humanserum. To each well added 40 μL of the serum containing LPS. Cover withfoil and mix by inverting the block. If the blood fails to clot or showsextensive hemolysis, the LPS can be prepared in the same way using humanType AB serum (Cambrex).

After a 4-hour incubation at 37° C. the blocks are centrifuged at 1000×gto fully pellet the RBC.

The plasma is harvested and placed into a polypropylene 96 well plateand then stored at 4° C.

Samples are tested for TNF or other cytokines as per the manufacturer'sinstructions (TNF ELISA; R&D Systems, Cat #DY210). For TNF the samplesare diluted 1:20 with PBS+1% BSA.

Human PBMC TNF Synthesis Assay

Human PBMC were obtained by from a healthy human donor leuko pak.Typically, all donors were pre-screened for STD, HIV and have been givena physical by a physician. One to two ampoules were thawed at 37° C. andthen resuspended in 45 to 85 mL of RPMI+10% FBS to a final concentrationof 2.5×10⁶ cells/mL. Resuspended cells were filtered to remove any largeclumps. (BD Falcon Cell strainer, 70 μM. Cat #352350) 00 μL of cellswere added to each well of a 96-well tissue culture plate (CorningCostar #3595) and equilibrate in a humidified 5% CO₂ incubator at 37° C.for 30 minutes.

Prepare compounds for addition to cells: All compounds were DMSO stocksat 20 mM. Working stock for cell testing were prepared as 4×concentrated material in RPMI+10% FBS. Compounds were added to apolypropylene block and then diluted with the appropriate amount ofRPMI+10% FBS. Typically 2.6 μL of 20 mM stock was diluted to 650 μLfinal volume to yield an 80 μM starting concentration in the stocks.RPMI was added to DMSO stock by pipetting into the tube or block. Thecompounds were then serial diluted with RPMI+10% FBS. The final DMSOconcentrations were not greater than 0.5% in the assay.

Compounds were added to the cells as 4× working stock. 50 μL of compoundwas prepared above is added to the cells and re-equilibrated in ahumidified 5% CO₂ incubator at 37° C. for 30 minutes.

A stock of LPS was prepared using RPMI+10% FBS. To prepare the stock,2.8 μL of LPS (5 mg/mL; Sigma Cat #L3024) was added to 35 mL of media.50 μL of the LPS in media was added to the cells. Control cultureswithout LPS received only RPMI+10% FBS. After an overnight incubation at37° C., the cell culture supernatant was harvested and stored in apolypropylene 96 well plate.

Samples were tested for TNF as per the manufacturer's instructions (TNFELISA; R&D Systems, Cat #DY210). The samples were serially diluted withPBS+1% BSA to the appropriate dilution condition for the cell prep,typically a 1:4 or 1:6 dilution.

TNF inhibition for select compounds of the present invention is shown inthe table below:

Ex. TNF Ex. TNF Ex. TNF # inhib. (μM) # inhib. (μM) # inhib. (μM) 1 0.184 14.2 5 0.27 10 0.41 16 0.68 17 0.39 18 1.1 28 3.5 29 0.92 30 2.5 322.6 33 20 34 3.6 37 20 38 0.45 39 1.3 41 0.5 45 0.2 46 1 47 0.9 48 1.451 0.03 54 0.006 55 0.163 72 0.004 73 0.009 75 0.006 78 0.001 83 0.00184 0.002 87 0.002 88 0.001 89 0.005 90 0.002 91 0.001 92 0.0024 930.0004 94 0.0004 95 0.001 96 0.0004 97 0.0006 98 0.0003 99 0.02 1000.0009 103 0.001 106 0.004 107 0.001 108 0.45 109 0.008 110 0.123 1110.074 112 0.28 113 0.0003 114 0.031 115 0.029 116 0.002 117 1.0 1180.029 120 0.24 124 50 126 0.002 127 0.006Preparation of Human Peripheral Blood Mononuclear Cells from Donor LeukoPaks

Human donor leukopaks are obtained from normal, healthy human donors(Analytical Biological Services). Donor leukopaks are received byovernight delivery. All blood paks are shipped wrapped in cold packs andabsorbent packing material.

The donor leukopak is opened aseptically in a hood using a scalpel ornew razor blade cleaned with alcohol.

The donor cells are decanted into a 180 ml conical polypropylene tube(Nalgene Nunc #CS48; VWR#21020-500 or BD tube (VWR #21008-943).Typically a donor leukopak will yield 70-90 mL of cells.

The donation bag is backwashed with 50 mL of sterile Hanks BalancedSalts Solution (HBSS; Invitrogen w/o phenol red). Decant into cells.

Bring cell suspension to a final volume of 180 mL with HBSS at roomtemperature.

Add 30 mL of cell suspension to each of 6×50 mL conical tubes (Corning).

Underlay the cell suspension with 15 mL of Histopaque (Histopaque 1.077g/mL; Sigma 10771) brought to room temperature. Use a 50 mL syringe andan 18 GA 6 inch spinal needle (BD #408360; 1.2 mM×152 mM; 18 GA) tounderlay the histopaque taking care not to disturb the interface.

Centrifuge the tubes for 30 minutes at 400×g at room temperature. Thebrake should be turned off on the centrifuge.

Aspirate the media from the tubes taking care not to disturb the cellsat the interface. Remove PBMC at the interface by aspiration taking carenot to disturb the residual erythrocyte pellet.

Pool cells and resuspend to 200 mL. Split into 4×50 mL polypropylenetubes and centrifuge at 400×g for 10 minutes. Aspirate supernatant andresuspend cells in 100 mL of HBSS. Repeat centrifuge wash step 2×. Finalpellets are resuspended in 50 mL of HBSS and filtered to remove anylarge clumps. (BD Falcon Cell strainer, 70

M. Cat #352350)

Cells are counted using a hemacytometer. Dilute 0.5 mL of cells in 9.5mL of HBSS and count. Estimate cell number.

Pellet cells at 400×g for 10 minutes. Resuspend cells in fetal bovineserum (heat inactivated; Invitrogen) containing 6% DMSO to a finalconcentration of 1×10⁸ cells/mL.

Aliquot cells to cryovials, 1 mL per vial.

Place cells into cell freezer blocks and place into −80° C. freezerovernight.

Remove cells from freezer and place cells into liquid nitrogen storage.

Prior to use of cells in compound profiling assays, determine theability of the cells to respond to LPS (lipopolysaccharides fromEscherichia coli 0111:B4; Sigma Cat#L3024, 5 mg resuspended in 1 mL ofRPMI) and S100b (bovine S100b; Calbiochem, Cat#559290, resuspended in 1mL of RPMI+10% FBS).

Thaw one ampoule of cells and resuspend in 40 mL of RPMI (ATCC; Cat#30-2001)+10% fetal bovine serum to a final concentration of 2.5×10⁶cells/mL.

Add 100 μL of cells to each well of a 96-well tissue culture plate(Corning Costar #3595) and equilibrate in a humidified 5% CO₂ incubatorat 37° C. for 30 minutes.

Add 100 μL of RPMI+10% FBS with or without LPS (100 ng/mL) or S100 (20ug/mL).

After an overnight incubation at 37° C., harvest the cell culturesupernatant and store in a polypropylene 96 well plate at 4° C.

Samples are tested for TNF or other cytokines as per the manufacturersinstructions (TNF ELISA; R&D Systems, Cat #DY210). The samples areserially diluted with PBS+1% BSA to find the proper dilution conditionfor the cell prep.

PDE4 Inhibition Assay

A TR-FRET—based phosphodiesterase assay kit from “Molecular Devices” wasused to test whether these compounds were indeed direct inhibitors ofthe PDE4 enzyme. Using Roflumilast as a positive control, compounds ofthe present invention were tested against fluorescein-labeled cAMPsubstrate. The principle of the assay is based on the binding of anucleotide monophosphate generated upon cAMP conversion to 5′ AMP by PDEto an “IMAP binding reagent”, which in turn is also ligated to aseparate complex carrying terbium (Tb)-donor molecule. Proximity of thefluoreceinated 5′ AMP to the Tb donor generates Fluorescence ResonanceEnergy Transfer. A PDE inhibitor will reduce the conversion of cAMP to5′ AMP, thus reducing monophosphate that can bind to the IMAP bindingreagent and reduce the resulting FRET signal.

The following procedures were taken to obtain the experimental datadescribed below:

The assay buffer was prepared by diluting 5× supplied Tween-based bufferin 1:5 in water to make 1× buffer. Add desired additive to buffer (DTTor MnCl₂). In a separate 96-well polypropylene plate, compound dilutionswere prepared in assay buffer. Separate microcentrifuge tubes wereprepared of PDE4B and PDE4D according to assay template—in assay buffer.The tubes were kept on ice. The enzyme concentration shown on thetemplate were diluted by 1:4. FAM-cAMP substrate solution was preparedaccording to assay template. 5 μl compound was transferred frompolypropylene plate into black 384-well plate. This plate wascentrifuged briefly to make sure all 5 μl is on the bottom. Up to 80 μlof prepared PDE4B enzyme solution was transferred into alternate wellson row ‘N’ of 384-well plate starting from cell N1. Up to 80 μl ofprepared PDE4D enzyme solution was transferred into alternate wells onrow ‘O’ of 384-well plate, starting from cell O2. cAMP substratesolution was transferred into bottom row of separate 96-well plate. 5 μlof enzyme solution from the “reservoir” row (N or O) was transferred toeach of the wells containing compounds, per layout map. Next, 10 μl ofcAMP substrate was transferred to these wells. The order ofsubstrate-first or enzyme-first can be switched depending on what isoptimal. The final cAMP concentration was 100 nM in the reaction. 20 μlof assay buffer was pipetted into 4 separate wells—these are the blanks.The plate was sealed with an aluminum strip and incubated at 30° C. for90 minutes. A TR-FRET solution was prepared. 4 ml of 1× IMAP Buffer Awas added to 6 ml of IMAP Buffer B. 25 μl (1/800 of 20 ml) of bindingbeads was added to this and mix by inverting. 60 μl of this mixture waspipetted into 2 of the wells containing the “blank” assay buffer. Next,49.7 μl (1/400 of remaining volume) of Tb donor solution was added tothe remaining TR-FRET solution and mixed by inverting. 60 μl of thissolution was pipetted into remaining 2 “blank” assay buffer-containingwells. The TR-FRET solution was poured into a pipette boat and amultichannel pipette was used to drop 60 μl of solution into all assaywells. The wells were covered with a foil strip and incubated for atleast 3 hours or overnight protected from light (e.g. in a drawer) atroom temperature. The plate was read on the Envision Reader: Emission 1:520/Emission 2: 486/Exc: 340. Mirror: Umbelliferone (UV).

The data below was obtained by the procedure described above:

Ex. PDE4B2 PDE4D Ex. PDE4B2 PDE4D # IC50 (nM) IC50 (nM) # IC50 (nM) IC50(nM) 30 17000 25000 32 25000 25000 33 12400 25000 51 126 616 54 78 33884 513 827 92 68 151 103 17 55 115 49 163 118 80 206 121 26 59 122 39 80127 25 73

The specific pharmacological responses observed may vary according toand depending on the particular active compound selected or whetherthere are present pharmaceutical carriers, as well as the type offormulation and mode of administration employed, and such expectedvariations or differences in the results are contemplated in accordancewith practice of the present invention.

Although specific embodiments of the present invention are hereinillustrated and described in detail, the invention is not limitedthereto. The above detailed descriptions are provided as exemplary ofthe present invention and should not be construed as constituting anylimitation of the invention. Modifications will be obvious to thoseskilled in the art, and all modifications that do not depart from thespirit of the invention are intended to be included with the scope ofthe appended claims.

We claim:
 1. A method of treating ulcerative colitis or Crohn's diseasecomprising administering to a human a compound, wherein the compound isselected from the group consisting of1-cyclopentyl-7-methoxy-3-(4-methyl-thiophen-2-yl)-1H-pyrimido[5,4-c]quinoline-2,4-dione,1-cyclopentyl-7-methoxy-3-m-tolyl-1H-pyrimido[5,4-c]quinoline-2,4-dione,3-(3-chloro-phenyl)-1-(1-methane-sulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]-quinoline-2,4-dione,trans-4-[3-(3-chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido-[5,4-c]quinolin-1-yl]-cyclohexanecarboxylicacid methyl ester,3-(3-chloro-phenyl)-1-((R)-1-methane-sulfonyl-pyrrolidin-3-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione,3-(3-chloro-phenyl)-1-((S)-1-methane-sulfonyl-pyrrolidin-3-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione,3-(3-chloro-phenyl)-1-(1-ethanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido-[5,4-c]quinoline-2,4-dione,andN-{4-[(S)-3-(3-chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-trans-cyclohexyl}-methanesulfonamide,or a pharmaceutically acceptable salt thereof.
 2. The method of claim 1,wherein the compound is1-cyclopentyl-7-methoxy-3-(4-methyl-thiophen-2-yl)-1H-pyrimido[5,4-c]quinoline-2,4-dioneor a pharmaceutically acceptable salt thereof.
 3. The method of claim 1,wherein the compound is1-cyclopentyl-7-methoxy-3-m-tolyl-1H-pyrimido[5,4-c]quinoline-2,4-dioneor a pharmaceutically acceptable salt thereof.
 4. The method of claim 1,wherein the compound is3-(3-chloro-phenyl)-1-(1-methane-sulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]-quinoline-2,4-dioneor a pharmaceutically acceptable salt thereof.
 5. The method of claim 1,wherein the compound istrans-4-[3-(3-chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido-[5,4-c]quinolin-1-yl]-cyclohexanecarboxylicacid methyl ester or a pharmaceutically acceptable salt thereof.
 6. Themethod of claim 1, wherein the compound is3-(3-chloro-phenyl)-1-((R)-1-methane-sulfonyl-pyrrolidin-3-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dioneor a pharmaceutically acceptable salt thereof.
 7. The method of claim 1,wherein the compound is3-(3-chloro-phenyl)-1-((S)-1-methane-sulfonyl-pyrrolidin-3-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dioneor a pharmaceutically acceptable salt thereof.
 8. The method of claim 1,wherein the compound is3-(3-chloro-phenyl)-1-(1-ethanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido-[5,4-c]quinoline-2,4-dioneor a pharmaceutically acceptable salt thereof.
 9. The method of claim 1,wherein the compound isN-{4-[(S)-3-(3-chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido[5,4-c]quinolin-1-yl]-trans-cyclohexyl}-methanesulfonamideor a pharmaceutically acceptable salt thereof.
 10. The method of claim1, wherein the compound is administered orally.
 11. A method of treatingulcerative colitis or Crohn's disease comprising administering to ahuman a compound, wherein the compound istrans-4-[3-(3-chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido-[5,4-c]quinolin-1-yl]-cyclohexanecarboxylicacid or a pharmaceutically acceptable salt thereof.
 12. The method ofclaim 11, wherein the compound istrans-4-[3-(3-chloro-phenyl)-7-methoxy-2,4-dioxo-3,4-dihydro-2H-pyrimido-[5,4-c]quinolin-1-yl]-cyclohexanecarboxylicacid.
 13. The method of claim 11, wherein the compound is administeredorally.
 14. A method of treating ulcerative colitis or Crohn's diseasecomprising administering to a human a compound, wherein the compound is3-benzo[1,3]dioxol-5-yl-1-(1-methanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dioneor a pharmaceutically acceptable salt thereof.
 15. The method of claim14, wherein the compound is3-benzo[1,3]dioxol-5-yl-1-(1-methanesulfonyl-piperidin-4-yl)-7-methoxy-1H-pyrimido[5,4-c]quinoline-2,4-dione.16. The method of claim 14, wherein the compound is administered orally.